Coumarin-amide derivatives and its preparation, said drug composition and its use

ABSTRACT

The present invention relates to novel coumarin derivatives, their carboxamides, pharmaceutical compositions containing them and their uses as drugs for kidney protection, treating drugs of hypertension, cardio-cerebrovascular diseases, non-insulin dependent diabetes, tumor, pre-cancerous lesion, and edemas.

TECHNICAL FIELD

The present invention relates to novel coumarins, their carboxamidederivatives, the preparation method thereof and the pharmaceuticalcompositions containing them, and their use as medicaments for kidneyprotection, as well as for the treatment of hypertension,cardio-cerebrovascular diseases, non-insulin dependent diabetes (NIDD),tumor, preneoplastic lesions, and edemas.

BACKGROUND ART

In 1990 the German Federal Institute of Drugs and Medicinnal Products(BfArM) published a monograph on Meliloti Herba, in which the use ofmelilot is indicated for symptoms and signs in chronic venousinsufficiency like pains; adjuvant treatment of thrombophlebitis andlymphostasis. Scheel et al. (Microbiol Toxins 8: 47-66, 1972) reportedthat coumarins have anti-bacterial, anti-viral and anti-tumor effects.Kovach et al (Arzeim-Forsch/Drug Res 20: 1630-33, 1970) proved thatcoumarins can increase blood flow and improve myocardial ischemia.Casley-Smith, (Vasomed 6: 232-4, 1994), Gaffney (J Pathol 133: 229-42,1981), Piller (Br J Exp Pathol 59: 319-26, 1978), and Knight (Clin Sci77: 69-76, 1989) showed that coumarins have effects of endothelialsystem protection and lymph-circulation promotion, etc. Nair et al.(Carcinogenesis 12 (1): 65-69, 1991) reported the anticancer activity ofcoumarins compounds. Ishizuka et al. (U.S. Pat. No. 5,096,924) provedthat substituted coumarins have anticancer activities. Marshall et al.(J. Biol. Resp. Mod. 8: 62, 1989) reported that coumarins haveimmuno-enhancing effects, such as improving the antitumor abilities byremarkably raising monocytes of patients suffering from cancer.Preuss-Ueberschar et al. (Drug Res. 34: 1305-1313, 1984) showed that thecoumarins are non-teratogenitic. Takagaki, Hidetsugu et al. (EP 0, 796,854 A1, 1997) disclosed that 3-, 4-, or 7-hydroxy or alkoxy substitutedcoumarins' effects in treating heart diseases. Markal et al. (WO 98/25,608, 1998) disclosed that substituted 4-arylcoumarins can be used totreat viral infections, such as herpes zoster or herpes simplex.Trkovnik et al. (WO 99/21550, 1999) reported that4-methyl-7-hydroxycoumarin can be used to treat or preventnephrocirrhosis, pancreatitis, and disorders in bladder or alimentarytracts. Takagaki et al. (Jpn. Kokai Tokkyo Koho JP 07277972, 1995)reported that coumarin derivatives can inhibit rat diabetes induced bystreptozotocin. Scott et al. (U.S. Pat. No. 5,723,476, 1998) disclosedthat 3-carboxamide-4-hydroxy coumarin compounds are effective to thenon-insulin dependent diabetes models. Han, Xingmei et al. (ZhongguoYaolixue Tongbao, 15(4): 332-5, 1999) reported that6,7-dimethoxycoumarin is effective to acute renal failure induced byendotoxins. Allen et al. (PCT Int Appl WO 2001 019396 A1 2001) reportedthat the TGF-β1 antagonists may be used for the treatment or prophylaxisof chronic nephritis.

In our research, a series of coumarin derivatives were synthesized andtheir biological activities were evaluated. For example, Xiao-long Huanget al reported substituted 3-acetyl- and 3-glyoxal-coumarin derivativespossessing good anti-mutagenic effects (Yaoxue Xuebao 31(6): 431-436,1996; ibid 31(7): 509-516, 1996). Shi-ping Xu et al discovered that thecoumarin retinoids show potent differentiation inducing, anti-mutagenic,and anti-carcinogenic effects (Chinese patent application No.97116602.1, CN1207392A). Song Xu et al.'s study on 6- or7-substitutedstyryl-coumarins, 4-styryl-coumarins and 4-, 6- or7-substitutedphenyliminomethylene-coumarins show anticancer effects(Yaoxue Xuebao 35(2): 103-107, 2000; ibid 36(4): 269-273, 2001; ibid37(2): 113-116, 2002).

Following that, upon our continued research works on coumarinscompounds, a series of novel coumarins and coumarin carboxamides weresynthesized. And we have found that these coumarins carboxamidecompounds possess potent inhibition effects on transforming growthfactor β1 (TGF-β1), which has not been reported before. The TGF-β1inhibitors may be used for the treatment of chronic renal dysfunctionsand diabetic renal dysfunctions. Meanwhile, it can also significantlydecrease angiotensin II (AngII). Therefore, the compounds of presentinvention, may be used in the drugs for the treatment of chronic renalfailure, nephritis, hypertension, even cirrhosis of liver and pulmonaryfibrosis. For example, Allen et al. (PCT Int Appl WO 2001 019396 A12001) reported that the TGF-β antagonists may be used for the treatmentor prophylaxis of chronic nephritis.

Renal insufficiencies, particularly chronic renal failure, are theresults of kidney injuries with various pathogenesis and progressivedeterioration. Among the primary nephropathies, the most common is thechronic glomerulonephritis, and tubolointerstitial nephritic comes thesecond. Among the secondary nephropathies, diabetic nephropathy holdsthe first position. At present, diabetic nephropathy holds about 27% ofthe origin of chronic renal failures, and is still increasing;hypertensive nephropathy comes the second, about 22.7% and theglomerulonephritis comes next about 21.2% and all other pathogenesisoccupy 26.6% in the origin of chronic renal insufficiencies. Being acommon disease per se, nephropathies, no wonder what pathogenesis, orbeing immune or non-immune mechanism, unless promptly treated, may beresult in chronic renal insufficiency and irreversible chronic renalinjuries.

Upon the researches of the field, it shows that transforming growthfactor-β1 (TGF-β1) has a close relationship with chronic renalinsufficiencies caused by various pathogenesis. Four hours afternephrectomy, TGF-β1 began to increase and the renin-angiotensin systemwas consequently influenced. Continuously rising of TGF-β1 andover-expression will result in inhibiting the degradation of theextracellular matrix, promoting the matrix integration, and also relatesto the proteinuria from renal insufficiency, as well as matrix fibrosis.Therefore, glomerular sclerosis and interstitial fibrosis have directrelationship with TGF-β1, and renin-angiotensin system and TGF-β1 arethe two critical factors related to chronic renal insufficiencies.Moreover, as the inhibition of the former has close relationship withthe decrease of TGF-β1 producing, this suggests that the increase ofTGF-β1 might be an important pathogenesis of kidney injuries to theend-stage renal insufficiencies, and would be a new target for thescreening of new anti-renal failure drugs.

Coumarin compounds possess extensive biological activities, however, ithaven't been reported in the literatures that these compounds can beused for the treatment of chronic renal failures. The present compoundsare a novel type compounds and can remarkably inhibit TGF-β1, which isan important pathogenesis of kidney injuries to the end-stage renalfailure. It has been proved that, the compound of present invention showsatisfied effects on treating renal insufficiencies.

Renal insufficiencies, especially chronic renal insufficiencies, arechronic diseases that are hard to be cured. With the continuouslyincrease of diabetes and hypertension, the incidence of renalinsufficiencies becomes more and more, and has no effective drugs orother methods for the treatment or prophylaxis up to now. Therefore, theobject of the present invention is to develop new drug for the treatmentof renal insufficiencies.

SUMMARY OF THE INVENTION

An object of the present invention is to overcome one or moredeficiencies of the prior arts, and to provide a new coumarin, inparticular to provide low toxic carboxamide derivatives thereof.

One another object of the present invention is to provide a preparationmethod of the coumarin carboxamide derivatives.

One aspect of the present invention relates to pharmaceuticalcompositions, which comprises a compound of general formula (I) or anisomer thereof as the active ingredient, and pharmaceutically acceptablecarriers.

A further object of the present invention is to provide use of the novelcoumarin carboxamide derivatives or the compositions thereof for the asTGF-β1 and angiotensin II (AngII) inhibitors.

A still further object of the present invention is to provide use of thenovel coumarin carboxamide derivatives or the compositions thereof forthe preparation of the medicaments for the treatment of kidney disorders(such as various chronic nephritis, diabetic and hypertensive renalinsufficiency), non-insulin dependent diabetes, cardio-cerebrovasculardiseases and hypertension.

Specifically, the first aspect of the present invention relates to thecompounds of the general formula (I)

Wherein,

R³ is selected from the group consisting of H, carboxyl,alkyloxycarbonyl, 5′-(phenyloxadiazol-2′)-yl,5′-(pyridyl-4″-oxadizol-2′)-yl,

CONHR₉, wherein R₉ is selected from the group consisting of C₂-C₈ fattyacid, benzoxamido, isonicotinamido, un-substituted or mono- ormulti-substituted phenyl wherein the substituent may be hydroxyl, C₁-C₈alkoxyl, CF₃, carboxyl, alkyloxycarbonyl, OCH₂CO₂H, NO₂, halogen, SO₃H,SO₂NHR₁₁, wherein R₁₁ is selected from the group consisting of hydrogen,amidino, 2″-thiazolyl, 3″-(5″-methylisooxazolyl), 2″-pyrimidinyl,2″-(4″,6″-dimethylpyrimidinyl), 4″-(5″,6″-dimethoxypyrimidinyl);

R₄ is selected from the group consisting of hydrogen, CONHR₁₀, whereinR₁₀ is selected from the group consisting of C₂-C₈ fatty acid,benzoxamido, isonicotiniamido, un-substituted, mono- ormulti-substituted phenyl wherein the substituent may be hydroxyl, C₁-C₈alkoxyl, CF₃, carboxyl, alkoxycarbonyl, OCH₂CO₂H, NO₂, halogen, SO₃H,SO₂NHR₁₂, wherein R₁₂ is selected from the group consisting of H,amidino, 2″-thiazolyl, 3″-(5″-methylisooxazolyl), 2″-pyrimidinyl,2″-(4″,6″-dimethyl-pyrimidinyl), 4″-(5″,6″-dimethoxy pyrimidinyl);

R₅ is selected from the group consisting of H, C₁-C₄ alkyl;

R₆ is selected from the group consisting of H, C₁-C₁₂ alkyl, halogen,NO₂, CONHR₁₃, wherein R₁₃ is substituted phenyl;

R₇ is selected from the group consisting of H, hydroxyl, C₁-C₄ alkyl oralkoxyl, carboxylalkylenoxyl, OCH₂CONHR₁₄, wherein R₁₄ is selected fromthe group consisting of un-substituted, mono- or multi-substitutedphenyl wherein the substituent may be hydroxyl, OCH₃, CF₃, CO₂H,CO₂C₂H₅, NO₂;

R₈ is selected from the group consisting of H, C₁-C₄ alkyl or alkoxyl,NO₂;

In order to achieve the object of the present invention, preferablecompounds include, but are not limited to the following compounds:

R₃ is selected from the group consisting of H, COOH, CO₂C₂H₅,5′-(phenyloxadiazol-2′)-yl, 5′-(pyridyl-4″-oxadizol-2′)-yl,

CONHR₉, wherein R₉ is n-butyric acid, o-, m-, p-phenol, o-, m-,p-carboxyl-phenyl, o-, m-, p-alkyloxycarbophenyl, methoxylphenyl,3′-hydroxy-4′-carboxyphenyl, 3′-salicylyl, 4′-salicylyl, m-CF₃-phenyl,3′-CF₃-4′-NO₂-phenyl, 2′-CO₂H-4′-I-phenyl, isonicotinamido, benzoxamido,3′-carboxy-methylenoxyphenyl, 4′-amidosulfonylphenyl,4′-guanidinosulfonylphenyl, 4′-(2″-thiazolamidosulfonyl)phenyl,4′-(5″-methylisooxazolyl-3″-amidosulfonyl)phenyl,4′-(pyrimidinyl-2″-amidosulfonyl)phenyl,4′-(4″,6″-dimethylpyrimidinyl-2″-amidosulfonyl)phenyl,4′-(5″,6″-dimethoxypyrimidinyl-4″-amidosulfonyl)phenyl;

R₄ is selected from the group consisting of H, CONHR₁₀, wherein R₁₀ isselected from the group consisting of H, 4′-CO₂H-phenyl,4′-CO₂C₂H₅phenyl, 3′-CF₃-phenyl;

R₅ is selected from the group consisting of H, CH₃;

R₆ is selected from the group consisting of H, C₂H₅, n-C₆H₁₃, NO₂, NH₂,Cl, Br, CONHR₁₃, wherein R₁₃ is selected from the group consisting ofcarboxyl- and alkoxycarbonyl-substituted phenyl;

R₇ is selected from the group consisting of H, OH, CH₃, OCH₃,OCH₂CONHR₁₄, wherein R₁₄ is selected from the group consisting ofphenyl, o-, m- and p-hydroxyphenol, o-, m- and p-carboxylphenyl, m- andp-ethoxycarbonylphenyl, m-CF₃-phenyl, m-CF₃-p-NO₂-phenyl, p-CH₃O-phenyl,4-salicylyl, 3-salicylyl;

R₈ is selected from the group consisting of H, CH₃, OCH₃, NO₂;

In order to complete the object of the present invention, preferablecompounds include, but are not limited to the compounds represented byfollowing general formula (Ia):

wherein R₄, R₅, R₆, R₇, R₈ are defined same as general formula

In order to complete the object of the present invention, preferablecompounds include, but are not limited to the compounds represented bythe following general formula (Ib):

wherein R₄, R₅, R₆, R₇, R₈, are same as defined in general formula (I),

R′₂ is selected from the group consisting of H, OH, CO₂H, etc;

R′₃ is selected from the group consisting of H, OH, CO₂H, CF₃, OCH₂CO₂H,etc;

R′₄ is selected from the group consisting of H, OH, CO₂H, CO₂Et, Iodo,NO₂, OCH₃, SO₃H, SO₂NH₂, SO₂NH(C═NH)NH₂,

etc;

R′₅ and R′₆ each is H;

In order to complete the object of the present invention, preferablecompounds include, but are not limited to the compounds of the followingtables 1 and 2:

Wherein R₄-R₈ groups are H except specified, R′₂-R′₆ groups are H exceptspecified

TABLE 1       No.       R₄—R₈

      MP ° C. 1 7-OCH₃ 4′-COOH >300   2 7-OCH₃ 3′-COOH >300   3 7-OCH₃2′-COOH >300   4 7-OCH₃ 2′-OH >300   5 7-OCH₃ 3′-OH 265 6 7-OCH₃4′-OH >300   7 7-OCH₃ 3′-OH, 4′-COOH  279d 8 7-OCH₃ 3′-COOH, 4′-OH  230d9 7-OCH₃ 2′-COOH, 4′-I >300   10 7-OCH₃ 4′-COOEt 247 11 7-OCH₃ 3′-CF₃218 12 7-OCH₃ 3′-CF₃ 4′-NO₂ >300   13 7-OCH₃ 4′-SO₂NH₂ >300   14 7-OCH₃4′-SO₂NH(C═NH)NH >300   15 7-OCH₃

>300   16 7-OCH₃

>300   17 7-OCH₃

298 18 7-OCH₃

300 19 7-OCH₃

 282d 20 7-OCH₃ 4′-OCH₃ 233 21 7-OCH₃ 4′-SO₃H 284 22 6-Et  7-OCH₃4′-COOH >300   23 6-Et  7-OCH₃ 3′-COOH 298 24 6-Et  7-OCH₃ 2′-COOH 29425 6-Et  7-OCH₃ 4′-OH 304 26 6-Et  7-OCH₃ 3′-OH, 4′-COOH 266 27 6-Et 7-OCH₃ 3′-COOH, 4′-OH 298 28 6-Et  7-OCH₃ 4′-COOEt 233 29 6-Et  7-OCH₃3′-CF₃ 224 30 6-Et  7-OCH₃ 3′-CF₃ 4′-NO₂ 234 31 6-Et  7-OCH₃4′-SO₂NH₂ >300   32 6-Et  7-OCH₃ 4′-SO₂NH(C═NH)NH 299 33 6-Et  7-OCH₃

>300   34 6-Et  7-OCH₃

>300   35 6-Et  7-OCH₃

278 36 6-Et  7-OCH₃

 260d 37 6-Et  7-OCH₃

>300   38 6-Et  7-OCH₃ 4′-OCH₃ 202 39 6-Et  7-OCH₃ 4′-SO₃H >300   407-OCH₃  8-CH₃ 4′-COOH >300   41 7-OCH₃  8-CH₃ 3′-COOH >300   42 7-OCH₃ 8-CH₃ 2′-COOH 264 43 7-OCH₃  8-CH₃ 3′-OH, 4′-COOH 279 44 7-OCH₃  8-CH₃3′-COOH, 4′-OH 290 45 7-OCH₃  8-CH₃ 2′-COOH, 4′-I 284 46 7-OCH₃  8-CH₃4′-COOEt 270 47 7-OCH₃  8-CH₃ 3′-CF₃ 258 48 7-OCH₃  8-CH₃ 3′-CF₃, 4′-NO₂252 49 7-OCH₃  8-CH₃ 4′-SO₂NH₂ 300 50 7-OCH₃  8-CH₃4′-SO₂NH(C═NH)NH >300   51 7-OCH₃  8-CH₃

>300   52 7-OCH₃  8-CH₃

277 53 7-OCH₃  8-CH₃

 220d 54 7-OCH₃  8-CH₃

286 55 7-OCH₃  8-CH₃

286 56 7-OCH₃  8-CH₃ 4′-OCH₃ 258 57 7-OCH₃  8-CH₃ 4′-SO₃H 286 58 7-OCH₃ 8-OCH₃ 4′-COOH 315 59 7-OCH₃  8-OCH₃ 3′-OH, 4′-COOH 264 60 7-OCH₃ 8-OCH₃ 3′-COOH, 4′-OH 264 61 7-OCH₃  8-OCH₃ 4′-COOEt 236 62 7-OCH₃ 8-OCH₃ 3′-CF₃ 243 63 7-OCH₃  8-OCH₃ 3′-CF₃, 4′-NO₂ 283 64 7-OCH₃ 8-OCH₃ 3′-OCH₂COOH 188 65 7-OCH₃  8-OCH₃ 4′-SO₂NH₂ 280 66 7-OCH₃ 8-OCH₃ 4′-SO₂NH(C═NH)NH 252 67 5-CH₃  7-OCH₃ 4′-COOH 299 68 5-CH₃ 7-OCH₃ 3′-COOH >300   69 5-CH₃  7-OCH₃ 2′-COOH >300   70 5-CH₃  7-OCH₃2′-OH 246 71 5-CH₃  7-OCH₃ 3′-OH 292 72 5-CH₃  7-OCH₃ 4′-OH 255 73 5-CH₃ 7-OCH₃ 3′-OH, 4′-COOH 284 74 5-CH₃  7-OCH₃ 3′-COOH  4′-OH >300   755-CH₃  7-OCH₃ 4′-COOEt 265 76 5-CH₃  7-OCH₃ 3′-CF₃ 221 77 5-CH₃  7-OCH₃3′-CF₃, 4′-NO₂ >300   78 5-CH₃  7-OCH₃ 4′-SO₂NH₂ 283 79 5-CH₃  7-OCH₃4′-SO₂NH(C═NH)NH 293 80 5-CH₃  7-OCH₃

288 81 5-CH₃  7-OCH₃

>300   82 5-CH₃  7-OCH₃

 274d 83 5-CH₃  7-OCH₃

268 84 5-CH₃  7-OCH₃

271 85 5-CH₃  7-OCH₃ 4′-OCH₃ 210 86 6-Cl  7-OCH₃ 4′-COOH >300   87 6-Cl 7-OCH₃ 3′-OH, 4′-COOH 253 88 6-Cl  7-OCH₃ 3′-COOH, 4′-OH >300   89 6-Cl 7-OCH₃ 4′-COOEt 294 90 6-Cl  7-OCH₃ 3′-CF₃ 282 91 6-Cl  7-OCH₃4′-SO₂NH₂ >300   92 6-Cl  7-OCH₃ 4′-SO₂NH(C═NH)NH 302 93 6-Cl  7-OCH₃

317 94 6-Br  7-OCH₃ 4′-COOH >300   95 6-Br  7-OCH₃ 2′-COOH 288 96 6-Br 7-OCH₃ 3′-OH, 4′-COOH 298 97 6-Br  7-OCH₃ 2′-COOH, 4′-I >300   98 6-Br 7-OCH₃ 4′-COOEt 298 99 6-Br  7-OCH₃ 3′-CF₃ 284 100 6-Br  7-OCH₃4′-SO₂NH₂ 298 101 6-Br  7-OCH₃ 4′-OCH₃ 262 102 6-nHex  7-OCH₃ 4′-COOH248 103 6-nHex  7-OCH₃ 2′-COOH 272 104 6-nHex  7-OCH₃ 3′-OH, 4′-COOH 268105 6-nHex  7-OCH₃ 2′-COOH, 4′-I 249 106 6-nHex  7-OCH₃ 4′-COOEt 160 1076-nHex  7-OCH₃ 3′-CF₃ 201 108 6-nHex  7-OCH₃ 4′-SO₂NH₂ 242 109 6-nHex 7-OCH₃ 4′-OCH₃ 170 110 6-NO₂  7-OCH₃  8-OCH₃ 4′-COOH >300   111 6-NO₂ 7-OCH₃  8-OCH₃ 3′-COOH 232 112 6-NO₂  7-OCH₃  8-OCH₃ 4′-OCH₃ 256 1136-NO₂  7-OCH₃  8-OCH₃ 3′-OH 160 114 6-NO₂  7-OCH₃  8-OCH₃ 2′-OH 217 1156-NO₂  7-OCH₃  8-OCH₃ 4′-COOEt 193 116 6-NO₂  7-OCH₃  8-OCH₃ 3′-OH,4′-COOH >300   117 6-NO₂  7-OCH₃  8-OCH₃ 3′-COOH, 4′-OH  266d 118 6-NO₂ 7-OCH₃  8-OCH₃ 3′-CF₃ 218 119 6-NO₂  7-OCH₃  8-OCH₃ 3′-CF₃, 4′-NO₂ 217120 6-NO₂  7-OCH₃  8-OCH₃ 4′-SO₂NH₂  290d 121 6-NO₂  7-OCH₃  8-OCH₃4′-SO₂NH(C═NH)NH 284 122 6-NO₂  7-OCH₃  8-OCH₃

 190d 123 6-NO₂  7-OCH₃  8-OCH₃

 220d 124 6-NO₂  7-OCH₃  8-OCH₃

 200d 125 6-C₂H₅, 7-OH  8-NO₂ 4′-COOH 234 126 6-C₂H₅, 7-OH  8-NO₂4′-OCH₃  218d 127 6-C₂H₅, 7-OH  8-NO₂ 3′-OH >300   128 6-C₂H₅, 7-OH 8-NO₂ 2′-OH  248d 129 6-C₂H₅, 7-OH  8-NO₂ 4′-COOEt 160 130 6-C₂H₅, 7-OH 8-NO₂ 3′-OH, 4′-COOH >300   131 6-C₂H₅, 7-OH  8-NO₂ 3′-COOH,4′-OH >300   132 6-C₂H₅, 7-OH  8-NO₂ 3′-CF₃ 169 133 6-C₂H₅, 7-OH  8-NO₂4′-SO₂NH₂  206d 134 6-C₂H₅, 7-OH  8-NO₂ 4′-SO₂NH(C═NH)NH 181 135 6-C₂H₅,7-OH  8-NO₂

>300   136 6-C₂H₅  7-OCH₃  8-NO₂ 4′-COOH 273 137 6-C₂H₅  7-OCH₃  8-NO₂4′-OH 252 138 6-C₂H₅  7-OCH₃  8-NO₂ 4′-OCH₃ 169 139 6-C₂H₅  7-OCH₃ 8-NO₂ 4′-COOEt 186 140 6-C₂H₅  7-OCH₃  8-NO₂ 4′-SO₂NH(C═NH)NH  206d 1416-NO₂, 7-OH, 8-CH₃ 4′-COOH >300   142 6-NO₂, 7-OH, 8-CH₃ 2′-COOH 227 1436-NO₂, 7-OH, 8-CH₃ 4′-OH >300   144 6-NO₂, 7-OH, 8-CH₃ 3′-OH >300   1456-NO₂, 7-OH, 8-CH₃ 2′-OH >300   146 6-NO₂, 7-OH, 8-CH₃ 4′-OCH₃ 254 1476-NO₂, 7-OH, 8-CH₃ 4′-COOEt >300   148 6-NO₂, 7-OH, 8-CH₃ 3′-OH, 4′-COOH271 149 6-NO₂, 7-OH, 8-CH₃ 3′-COOH, 4′-OH >300   150 6-NO₂, 7-OH, 8-CH₃3′-CF₃ 231 151 6-NO₂, 7-OH, 8-CH₃ 3′-CF₃, 4′-NO₂ 226 152 6-NO₂, 7-OH,8-CH₃ 4′-SO₂NH₂ >300   153 6-NO₂, 7-OH, 8-CH₃ 4′-SO₂NH(C═NH)CH₂ >300  154 6-NO₂, 7-OH, 8-CH₃

>300   155 6-NO₂, 7-OH, 8-CH₃

254 156 6-NO₂, 7-OH, 8-CH₃

>300   157 6-NO₂, 7-OH, 8-CH₃ 2′-COOH, 4′-I 262 158 6-NO₂, 7-OCH₃, 8-CH₃4′-COOH 301 159 6-NO₂, 7-OCH₃, 8-CH₃ 3′-COOH 280 160 6-NO₂ 7-OCH₃, 8-CH₃2′-COOH 282 161 6-NO₂, 7-OCH₃, 8-CH₃ 4′-OH >300   162 6-NO₂, 7-OCH₃,8-CH₃ 3′-OH 231 163 6-NO₂, 7-OCH₃, 8-CH₃ 2′-OH 285 164 6-NO₂, 7-OCH₃,8-CH₃ 4′-OCH₃ 209 165 6-NO₂, 7-OCH₃, 8-CH₃ 4′-COOEt 230 166 6-NO₂,7-OCH₃, 8-CH₃ 3′-OH, 4′-COOH 249 167 6-NO₂, 7-OCH₃, 8-CH₃ 3′-CF₃ 182 1686-NO₂, 7-OCH₃, 8-CH₃ 3′-CF₃, 4′-NO₂ 236 169 6-NO₂, 7-OCH₃, 8-CH₃4′-SO₂NH(C═NH)NH >300   170 6-NO₂, 7-OCH₃, 8-CH₃ 4′-SO₂NH₂ 301 1716-NO₂, 7-OCH₃, 8-CH₃

276 172 6-NO₂, 7-OCH₃, 8-CH₃

270 173 6-NO₂, 7-OCH₃, 8-CH₃

299 174 6-NO₂, 7-OH, 8-NO₂ 4′-COOH >300   175 6-NO₂, 7-OH, 8-NO₂ 4′-OH260 176 6-NO₂, 7-OH, 8-NO₂ 3′-OH >300   177 6-NO₂, 7-OH, 8-NO₂2′-OH >300   178 6-NO₂, 7-OH, 8-NO₂ 4′-OCH₃ >300   179 6-NO₂, 7-OH,8-NO₂ 4′-COOEt 281 180 6-NO₂, 7-OH, 8-NO₂ 3′-CF₃ 197 181 6-NO₂, 7-OH,8-NO₂ 4′-SO₂NH₂ >300   182 6-NO₂, 7-OH, 8-NO₂ 4′-SO₂NH(C═NH)NH 216 1836-NO₂, 7-OH, 8-NO₂

>300   184 6-NO₂, 7-OH, 8-NO₂

170 185 6-NO₂, 7-OH, 8-NO₂

>300   186 6-NO₂, 7-OH, 8-NO₂ 2′-COOH 285 187 6-NO₂, 7-OCH₃  8-NO₂ 4′-OH257 188 6-NO₂, 7-OCH₃  8-NO₂ 4′-COOEt 236 189 6-NO₂, 7-OCH₃  8-NO₂4′-OCH₃ 205 190 6-Cl  7-OH  8-NO₂ 4′-OCH₃ 285 191 6-Cl  7-OH  8-NO₂4′-SO₂NH(C═NH)NH  300d 192 6-Cl  7-OH  8-NO₂ 3′-OH  4′-COOH >300   1935-CH₃, 6-, 8-(NO₂)₂  7-OH 4′-COOH >300   194 5-CH₃6-, 8-(NO₂)₂, 7-OH3′-COOH 246 195 5-CH₃6-, 8-(NO₂)₂  7-OH 2′-COOH 214 196 6-CH₃6-,8-(NO₂)₂  7-OH 4′-OCH₃ 242 197 5-CH₃6-, 8-(NO₂)₂  7-OH 4′-COOEt 244 1985-CH₃6-, 8-(NO₂)₂  7-OH 4′-SO₂NH₂ 256 199 5-CH₃, 6-, 8-(NO₂)₂  7-OH4′-SO₂NH(C═NH)NH >300   200 5-CH₃6-, 8-(NO₂)₂  7-OH

>300   201 5-CH₃6-, 8-(NO₂)₂  7-OH

220 202 5-CH₃6-, 8-(NO₂)₂  7-OH

276 R₄—R₈ R₃ 203 7-OCH₃ CONH(CH)₃CO₂H 193 204 7-OCH₃

293 205 7-OCH₃

248 206 7-OCH₃

238 207 6-C₂H₅, 7-OCH₃ CONH(CH)₃CO₂H 226 208 6-C₂H₅, 7-OCH₃

293 209 6-C₂H₅, 7-OCH₃

196 210 5-CH₃, 7-OCH₃

248 211 5-CH₃, 7-OCH₃

176 212 5-CH₃, 7-OCH₃

240 213 7-OCH₃, 5-CH₃

254 214 7-OCH₃, 8-CH₃

254 215 7-OCH₃  8-CH₃

278 216 7-OCH₃  8-CH₃

270 217 6-Br  7-OCH₃

248 218 6-Br  7-OCH₃

>300   219 6-Br  7-OCH₃

295 220 6-n-C₆H₁₃  7-OCH₃

198 221 6-n-C₆H₁₃  7-OCH₃

196 222 6-n-C₆H₁₃  7-OCH₃

139 223 6-NO₂, 7-OH, 8-CH₃

>300   224 6-NO₂, 7-OCH₃, 8-CH₃

220 225 6-NO₂  7,8-(OCH₃)₂

199 226 6-NO₂  7,8-(OCH₃)₂

>300  

TABLE 2 No. R₃ R₄ R₅ R₆ R₇ R₈ MP ° C. 227 CO₂C₂H₅ H H NO₂ OCH₃ OCH₃ 191228 CO₂H H H NO₂ OCH₃ OCH₃ 188 229 CO₂C₂H₅ H H NO₂ OH CH₃ 210 230 CO₂H HH NO₂ OH CH₃ 251 231 CO₂C₂H₅ H H NH₂ OH CH₃ 203 232 CO₂H H H NO₂ OCH₃CH₃ 178 233 CO₂C₂H₅ H H C₂H₅ OH NO₂ 143 234 CO₂H H H C₂H₅ OH NO₂ 178 235CO₂C₂H₅ H H C₂H₅ OCH₃ NO₂ 140 236 CO₂H H H C₂H₅ OCH₃ NO₂ 176 237 CO₂C₂H₅H H NO₂ OH NO₂ 176 238 CO₂H H H NO₂ OH NO₂ 296 239 CO₂C₂H₅ H H NO₂ OCH₃NO₂ 152 240 CO₂H H H NO₂ OCH₃ NO₂ 246 241 CO₂C₂H₅ H H Cl OH NO₂ 195 242CO₂H H H Cl OH NO₂ >300 243 CO₂H H CH₃ NO₂ OH NO₂ 211 244 CO₂C₂H₅ H CH₃NO₂ OH NO₂ 104 245 H

H H CH₃ H 223 246 H

H H CH₃ H >300 247 H

H H CH₃ H 197 248 H CH₃ H H

H 230 249 H CH₃ H H

H 220 250 H CH₃ H H

H 240 251 H CH₃ H H

H 196 252 H CH₃ H H

H 304 253 H CH₃ H H

H >300 254 H CH₃ H H

H 296 255 H CH₃ H H

H 207 256 H CH₃ H H

H 157 257 H CH₃ H H

H 243 258 H CH₃ H H

H 301 259 H CH₃ H H

H 180 260 H CH₃ H H

H 215 261 H CH₃ H H

H 277 262 H CH₃ H H

CH₃ 216 263 H CH₃ H H

CH₃ 205 264 H CH₃ H H

CH₃ 260 265 R₃ = R₄ = R₅ = R₇ = R₈ = H

214 266 R₃ = R₄ = R₅ = R₇ = R₈ = H

300

In this description, the term “halogen” indicates Fluoro, Chloro, Bromoand Iodo. The terms “lower alkane”, “lower alkyl” mean linear orbranched alkanes and alkyls having 1-6 carbon atoms.

According to the present invention, the compounds may have isomers,generally the said “compounds of the present invention” includes isomersthereof.

The compounds of the present invention may contain cis-/trans-isomers ofa double bond, asymmetric center with S- and R-configurations, andinclude all the potential stereoscopic isomers and mixtures of two ormore isomers. In case that cis-/trans-isomers exist, the presentinvention also relates to the cis- and trans-isomer and their mixtures,and a pure isomer may be separated according to the conventional methodsor synthesized from stereo-selective reagents if necessary.

According to the embodiments of present invention, said compounds mayalso include the pharmaceutically acceptable salts and hydrate(s),esters, or pro-drugs thereof.

According to the present invention, it is also related to thepreparation methods of the compounds of the present invention, which areprepared via nitrating or bi-nitrating various substituted 3-esteryl or3-carboxy-coumarins, thus to obtain part of the target compounds ofpresent invention and meanwhile the intermediates for the targetcompounds; reacting the intermediated acids, 3-carboxy-substitutedvarious substituted coumarins, 4-carboxy-substituted various substitutedcoumarins, 6-carboxy-substituted various substituted coumarins and7-carboxy-substituted various substituted coumarins with correspondingsubstituted amines to achieve the target compounds.

The amidating reaction is carried out with corresponding reactants andcatalysts, and in the suitable solvents. Said reactants includephosphorus trichloride, phosphorus oxychloride, phosphoruspentachloride, thionyl chloride, 1,3-dicyclohexylcarbodiimide,dipyridylcarbonate (2-DPC), 1,3-diisopropylcarbodiimide (DIPC), and1-(3-dimethylamino-propyl)-3-ethylcarbodiimide (EDCI), etc. Preferablereactants are phosphorus pentachloride, phosphorus trichloride andthionyl chloride, more preferable reactants are phosphorus pentachlorideand thionyl chloride. Catalytic agents for the preparation of thecompounds of present invention include tertiary amines, pyridine,4-dimethylaminopyridine and 4-pyrrolidylpyridine, preferably tertiaryamines and pyridine, more preferably pyridine. The reaction is carriedout in a suitable solvent or the above condensation agent, such asanhydrous aprotic solvent, dimethylsulfoxide (DMSO), toluene,dichloromethane, 1,2-dichloroethane, ethylene glycol dimethyl ether,tetrahydrofuran and N,N-dimethylformamide (DMF), preferably toluene,DMSO and DMF, more preferably toluene and DMF.

The reaction temperature is 10˜110° C., preferably 20˜90° C., morepreferably 30˜80° C., particularly preferably 50˜70° C.

The synthetic routes are specifically explained in the following routesIIa, IIb, IIc, IId, IIe and IIf.

Thus, the present invention also relates to a pharmaceutical compositioncontaining a compound of present invention as active ingredient andconventionally pharmaceutical excipients or auxiliaries. Generally thecomposition may comprise the compound of present invention from 0.1 to95% by weight.

The composition of present invention may be prepared by the commonmethods according to the art. For this purpose, the compounds of thepresent invention may be combined with one or more solid, semi-solid orliquid excipients and/or auxiliaries, to prepare a suitableadministration form or dosage forms for human or animal use.

The compounds of the present invention or compositions containing themmay be administered in the unit dosage forms and the administrationroutes, which can be oral or parenteral, such as oral, intramusculary,subcutaneously, nasal, oral mucosa, transdermal, intraperitoneal orrectal, etc.

The compounds of the present invention or compositions containing themmay be administered via injection, including intravenous, intramuscular,subcutaneous, intradermal, and acupoint etc.

The administration forms may be liquid or solid forms. For example theliquid forms may be solutions, colloids, fine particles, emulsion,suspensions and the like. Other forms may be such as tablets, capsules,sprays, dripping pills, aerosols, pills, powders, solutions,suspensions, emulsions, granules, suppositorys, freeze-dried powder forinjections and the like.

The present compounds can be prepared as normal preparations, orsustained-release, controlled-release or targeted preparations andvarious fine particle delivery systems.

In order to prepare unit administration forms to tablets, carriers wellknown in the art can be used. Carriers such as, diluents and absorbents,such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose,sodium chloride, glucose, urea, calcium carbonate, kaolin,microcrystallinecellulose, aluminum silicate and the like; moist andbinding agents for example water, glycerol, polyethyleneglycol, ethanol,propanol, starch paste, dextrin, syrups, honey, glucose solution, Arabiagum, gelatin, sodium carboxymethylcelluloses, lacta, methylcellulose,potassium phosphate, polyvinylpyrrolidone and the like; disintegrants,for example dry starch, alginates, agar powders, laminarin, sodiumhydrogencarbonate-citric acid, calcium carbonate,polyoxyethylene-sorbitol fatty acid esters, sodium lauryl sulfonate,methylcellulose, ethylcellulose and the like; disintegrant inhibitors,such as sucrose, glycerol tristearate, cocoa butter, hydrogenated oiland the like; absorb-promotors, for example quaternary ammonium salts,sodium lauryl sulfate and the like; lubricants, for example talc,silica, corn starch, stearates, boric acid, liquid paraffin,polyethyleneglycol and the like. The tablets may be further coated, forexample sugarcoating, film coating, enteric coating, or two ormulti-layered tablets.

To prepare pills from the administration units, carriers well known inthe art can be used. Carriers such as, diluents and absorbents, such asglucose, lactose, starch, cocoa butter, hydrogenated vegetable oils,polyvinylpyrrolidone, Gelucire, kaolin, talc and the like; bindingagents such as arabia gum, tragacanth, gelatin, ethanol, honey,liquid-sugars, rice pastes or flour pastes and the like; disintegrantssuch as agar powder, dry starch, alginates, sodium lauryl sulfonate,methylcellulose, ethylcellulose and etc.

To prepare capsules from the administration units, the compounds ofpresent invention may be mixed with the above carrier(s), and the soobtained mixtures are packaged into hard or soft capsules.Alternatively, the present compounds may also be prepared intomicrocapsules, and can be used as suspension in a hydrous media, orpackaged into hard capsules or injections.

To prepare the injection dosage forms, the compounds of presentinvention may be formulated into solutions, suspensions, emulsions,freeze dried powders for injections. Such formulations may be hydrous oranhydrous, which may contain one or two or more pharmaceuticallyacceptable carrier(s), diluents, preservatives, surfactants ordispersing agents. For example, diluents are selected from water,ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxyisostearylalcohol, polyoxyisostearyl alcohol, polyoxyethylene-sorbitol fatty acidesters and the like. In addition, to prepare the isotonic injections,sodium chloride, glucose or glycerol can be added to the injectionsolution, further, solubilizing agents, buffering agents, pH-modulatorsand the like can also be added.

Also, if desired, coloring agents, preservatives, perfumes, correctants,sweetening agents and the like, can also be added to the pharmaceuticalpreparation.

For achieving administering purpose and enhancing treating effect, thecompounds or pharmaceutical compositions of the present invention may beadministered by any known methods. The administering dosage of thepresent invention may extensively be varied by depending on a number offactors, for example the seriousness of the diseases to be treated orprevented, sex, age, body weights, disposition and individualdifferences of the patients or animals, administering routes or numberof times, treating purposes, etc. In general, the effective dosages ofthe pharmaceutically active ingredients are known to those skilled inthe art, the real administering dose can be suitably adjusted based onthe exact dosage contained in the formulations to achieve the treatmentor prophylaxis purposes.

The daily suitable ranges of dosages of the compounds of presentinvention are within 0.001-150 mg/kg body weight, preferably 0.1-100mg/kg, more preferably 1-60 mg/kg, most preferably 2-30 mg/kg. Thesedoses can be administered in one single dose or divided into severaldoses, for example twice, three or four times per day, which depends onthe experiences of doctors and the other different therapeutic means.

The total dose of each treatment may be administered in one portion ordivided into multi portions, depending on the total dose. The compoundor the pharmaceutical compositions of present invention may be adoptedalone or in combination with other drugs, in the latter case, the dosemay also be adjusted.

The activities and effects of the compounds and/or compositions of thepresent invention may be determined via in vitro and in vivo tests, suchas TGF-β1 antagonism, treating of the renal insufficiencies and thelike, which are all known in the filed. In recent years, researches haveconfirmed that TGF-β1 is one of most critical factors resulting in theprogressive renal failure with glomerulosclerosis and interstitialfibrosis.

Pharmacological tests show that the compounds of the present inventionpossess effects of blocking the binding of TGF-β1 with the receptors andinhibiting the production of TGF-β1. Of all the 33 subject compounds in10 μg/ml doses, 11 compounds possess activities of exceeding 50%, 8compounds possess activities of exceeding 60%, 7 compounds possessactivities of exceeding 70%, 5 compounds possess activities of exceeding80%, and 4 compounds possess activities of exceeding 90%.

In the cell growth inhibition model of mink pulmonary epithelial cellsinduced by TGF-β1, of all the 5 subject compounds, 3 compounds possessactivities of exceeding 60%, 2 compounds possess activities of exceeding70%, and 1 compound possesses activity of exceeding 90%. Thus, thepresent compounds can be used for the treatment or prophylaxis ofchronic renal disorders, including: a) primary nephropathies, commonlysuch as, the chronic glomerulonephritis, interstitial nephritis, chronicpyelonephritis and the like; b) secondary nephropathies, commonly suchas, chronic diabetic nephropathy, hypertensive nephropathy, lupusnephropathy and the like; c) congenital and obstructive diseases such aspolycystic kidney, posterior urethral valve disorders, neurogenicbladder hyperplasia, prostatic hyperplasia, urinary lithiasis, etc.

Additional researches show that the compounds of the present inventioncan remarkably inhibit the effect of Ang II (P<0.01). As mentionedabove, TGF-β1 and the renin-angiotesin system are closely related withrenal insufficiencies having multi pathogenesis. TGF-β1 and therenin-angiotesin system are the two most critical factors in theprogressive deterioration of renal disorders and the inhibition of AngII has a close relationship with the reduction of TGF-β1. As Ang IIplays an important role in the onset of various types of hypertensions,the compounds of the present invention may be used for the treatment ofrenal hypertensions, diabetic hypertensions, peripheral vascularhypertensions and cardio-cerebrovascular diseases having the abovepathogenesis.

In vivo tests on the model of chronic renal failure induced by a 5/6nephrectomy in rats showes that, comparing with the positive controlsBenazepril and Losartan, the compounds of the present inventionsuperiors than Benazepril and corresponds to (slightly better than)Losartan on reducing the blood serum urea nitrogen (BUN) and creatinine(Cre), as well as inhibiting TGF-β1 and Ang II.

In the tests of the renal interstitial fibrosis model from unilateralureteral ligation in rats, the subject compound 149 is better thanBenazepril and corresponds to (slightly better than) Losartan, and thepathological results show that the compound of present invention arebetter than Benazepril and correspond to Losartan.

The tested compounds have low toxicities, under the dosages of 5 g/kgbody weights and 10 g/kg body weights, within a continuous observationof two weeks, no death in the subjected mice were observed and no otherabnormal expressions were found.

In Ames test of the subjected compound 149, the negative results wasobtained which shows no mutagenesis.

EXAMPLES

The various starting materials of the examples can be prepared via theordinary method of the field or the methods commonly known by theskilled artisans, which can be prepared via e.g. the following reactionroutes.

-   (1) 3-ethoxycarbonyl-7-hydroxycoumarin and    3-carboxy-7-methoxycoumarin

-   (2) 3-ethoxycarbonyl-6-chloro-7-hydroxycoumarin and    3-carboxy-6-chloro-7-methoxycoumarin

-   (3) 3-ethoxycarbonyl-6-ethyl-7-hydroxycoumarin and    3-carboxy-6-ethyl-7-methoxycoumarin

-   (4) 3-ethoxycarbonyl-6-hexyl-7-hydroxycoumarin and    3-carboxy-6-hexyl-7-methoxycoumarin

-   (5) 3-ethoxycarbonyl-6-bromo-7-hydroxycoumarin and    3-carboxy-6-bromo-7-methoxycoumarin

-   (6) 3-ethoxycarbonyl-7,8-dihydroxycoumarin and    3-carboxy-7,8-dimethoxycoumarin

-   (7) 3-ethoxycarbonyl-7-hydroxy-8-methylcoumarin and    3-carboxy-7-methoxy-8-methylcoumarin

-   (8) 3-ethoxycarbonyl-7-hydroxy-5-methylcoumarin and    3-carboxy-7-methoxy-5-methylcoumarin

-   (9) 3-ethoxycarbonyl-6-nitro-7-hydroxy-8-methylcoumarin and    3-carboxy-6-nitro-7-methoxy-8-methylcoumarin

-   (10) 3-ethoxycarbonyl-6-nitro-7,8-dihydroxycoumarin and    3-carboxy-6-nitro-7,8-dimethoxycoumarin

-   (11) 3-ethoxycarbonyl-5-methyl-6,8-dinitro-7-hydroxycoumarin and    3-carboxy-5-methyl-6,8-dinitro-7-hydroxy-coumarin

-   (12) 3-ethoxycarbonyl-5-methyl-6,8-dinitro-7-methoxycoumarin and    3-carboxy-5-methyl-6,8-dinitro-7-methoxy-coumarin

-   (13) 3-ethoxycarbonyl-5-methyl-6,8-dinitro-7-hydroxycoumarin and    3-carboxy-5-methyl-6,8-dinitro-7-hydroxy-coumarin

-   (14) 3-ethoxycarbonyl-6-ethyl-7-hydroxy-8-nitrocoumarin and    3-carboxy-6-ethyl-7-hydroxy-8-nitrocoumarin

-   (15) 3-ethoxycarbonyl-6-chloro-7-hydroxy-8-nitrocoumarin and    3-carboxy-6-chloro-7-hydroxy-8-nitrocoumarin

-   (16) 6-carboxycoumarin

-   (17) 4-carboxy-7-methylcoumarin

-   (18) 4-methyl-7-carboxymethoxycoumarin

-   (19) 4,8-dimethyl-7-carboxymethoxycoumarin

The following examples are intended to illustrate this invention,however these examples shall not be construed to limit the scope of theinvention.

Example 1 Synthesis of3-ethoxycarbonyl-6-chloro-7-hydroxy-8-nitro-coumarin (241)

2.75 g (10.2 mmol) of 3-ethoxycarbonyl-6-chloro-7-hydroxycoumarin wasadded into 10 ml of concentrated sulfuric acid, 1.74 g (20.4 mmol) ofconcentrated nitric acid was added in portions under the cooling ofice-salt bath, the reaction was monitored through thin-layerchromatography to confirm the completion, and ice was added to seize thereaction. The reaction mixture was then filtered and washed by water,dried to give 1.52 g of the title compound (241).

¹H-NMR 300 MHz (DMSO): 1.266 (t, 3H, CH₃), 4.232 (q, 2H, CH₂), 8.017 (s,1H, 5-H), 8.593 (s, 1H, 4-H)

Compounds 229-246 in the tables were prepared following the sameprocedure.

Example 2 Synthesis of3-ethoxycarbonyl-6-ethyl-7-hydroxy-8-nitro-coumarin (233)

Compound 233 was prepared following the preparation of compound 241,except that 3-ethoxycarbonyl-6-ethyl-7-hydroxy-coumarin was nitrated togive the title compound 233.

¹H-NMR 300 MHz (DMSO): 1.262 (t, 3H, 6-ethyl-CH₃), 1.401 (t, 3H,ester-CH₃), 2.753 (q, 2H, 6-ethyl-CH₂), 3.988 (s, 3H, 7-OCH₃), 4.408 (q,2H, ester-CH₂), 7.527 (s, 1H, 5-H), 8.479 (s, 1H, 4-H)

Example 3 Synthesis of 3-ethoxycarbonyl-6-nitro-7,8-dimethoxy-coumarin(227)

Compound 227 was prepared following the preparation of compound 241,except that 3-ethoxycarbonyl-7,8-dimethoxycoumarin was nitrated to givethe title compound 227.

¹H-NMR 300 MHz (DMSO): 1.397 (t, 3H, ester-CH₃), 4.063-4.118 (d, 6H,7,8-OCH₃), 4.423 (q, 2H, ester-CH₂), 7.757 (s, 1H, 5-H), 9.252 (s, 1H,4-H)

Example 4 Synthesis of 3-ethoxycarbonyl-6,8-dinitro-7-methoxy-coumarin(239)

Compound was prepared following the preparation of compound 241, exceptthat 3-ethoxycarbonyl-7-methoxy-coumarin was bis-nitrated to give thetitle compound 239.

¹H-NMR 300 MHz (DMSO): 1.290 (t, 3H, ester-CH₃), 4.011 (s, 3H, 7-OCH₃),4.292 (q, 2H, ester-CH₂), 8.873 (s, 1H, 4-H), 8.955 (s, 1H, 5-H)

Example 5 Synthesis of 3-ethoxycarbonyl-6,8-dinitro-7-hydroxy-coumarin(237)

Compound 237 was prepared following the preparation of compound 241,except that 3-ethoxycarbonyl-7-hydroxy-coumarin was bis-nitrated to givethe title compound 237.

¹H-NMR 300 MHz (DMSO): 1.237 (t, 3H, ester-CH₃), 4.196 (q, 2H,ester-CH₂), 8.399 (s, 1H, 4-H), 8.636 (s, 1H, 5-H)

Example 6 Synthesis of3-(3′-hydroxy-4′-carboxy-phenylamidocarbonyl)-6-ethyl-7-methoxy-coumarin(26)

248 mg (1 mmol) of 3-carboxy-6-ethyl-7-methoxycoumarin and 2 ml of SOCl₂was heated to complete the reaction. After that, SOCl₂ was removed and153 mg (1 mmol) of 4-aminosalicylic acid and 2 ml of pyridine was added.The mixture was heated to complete the reaction. The crude product waspurified with DMSO to give 140 mg of the title compound (26).

¹H-NMR 300 MHz (DMSO): 1.142 (t, 3H, CH₃), 2.569 (q, 2H, CH₂), 3.906 (s,3H, 7-OCH₃), 7.069 (d, 1H, 6′-H), 7.098 (s, 1H, 8-H), 7.509 (s, 1H,2′-H), 7.758 (d, 1H, 5-H), 8.856 (s, 1H, 4-H), 10.848 (s, 1H, CONH),11.399 (s, 1H, OH) Compounds 1-109, 204-206, 208, 209, 213, 214, 217,218, 220, 222-228 in table 1 and compounds 247-249 were preparedfollowing the same procedure.

Example 7 Synthesis of3-(3′-carboxy-4′-hydroxy-phenylamidocarbonyl)-6-ethyl-7-methoxy-coumarin(27)

Compound 27 was prepared following the preparation of compound 26,except that 4-aminosalicylic acid was replaced by 5-amino-salicylic acidto give the title compound 27.

¹H-NMR 500 MHz (DMSO): 1.162 (t, 3H, ethyl-CH₃), 2.602 (q, 2H,ethyl-CH₂), 3.937 (s, 3H, 7-OCH₃), 6.786 (d, 1H, 5′-H), 7.178 (s, 1H,6-H), 7.746 (d, 1H, 6′-H), 7.770 (s, 1H, 5-H), 8.239 (s, 1H, 2′-H),8.834 (s, 1H, 4-H), 10.583 (s, 1H, CONH)

Element analysis for: C₂₀H₁₇NO₇ Calculated (%): C, 62.66; H, 4.47; N,3.65. Found (%): C, 62.87; H, 4.49; N, 3.71.

Example 8 Synthesis of3-(m-carboxyphenylamidocarbonyl)-7-methoxycoumarin (2)

Compound 2 was prepared following the preparation of compound 26, exceptthat 3-carboxy-7-methoxy-coumarin reacted with m-aminobenzoic acid togive the title compound 2.

Element analysis for: C₁₈H₁₃NO₆.½H₂O Calculated (%): C, 62.07; H, 4.05;N, 4.02. Found (%): C, 62.72; H, 3.74; N, 4.55.

Example 9 Synthesis of3-(3′-hydroxy-4′-carboxyphenylamidocarbonyl)-7-methoxy coumarin (7)

Compound 7 was prepared following the preparation of compound 26, exceptthat 3-carboxy-7-methoxy-coumarin reacted with 4-amino-salicylic acid togive the title compound 7.

¹H-NMR 300 MHz (DMSO): 3.91 (s, 3H, 7-OCH₃), 7.08 (d, 1H, 6-H), 7.11 (s,1H, 6′-H), 7.53 (s, 1H, 2′-H), 7.77 (d, 1H, 5-11), 7.95 (d, 1H, 5′-H),8.91 (s, 1H, 4-H), 10.83 (s, 1H, CONH), 11.40 (br, 1H, OH)

Element analysis for: C₁₈H₁₃NO₇ Calculated (%): C, 60.85; H, 3.69; N,3.94. Found (%): C, 60.52; H, 3.59; N, 4.10.

Example 10 Synthesis of3-(3′-carboxy-4′-hydroxyphenylamidocarbonyl)-7-methoxy coumarin (8)

Compound 8 was prepared following the preparation of compound 26, exceptthat 3-carboxy-7-methoxy-coumarin reacted with 5-amino-salicylic acid togive the title compound 8.

¹H-NMR 300 MHz (DMSO): 3.906 (s, 3H, 7-OCH₃), 6.964 (d, 1H, 5′-H), 7.037(d, 1H, 6-H), 7.083 (s, 1H, 8-H), 7.745 (d, 1H, 6′-H), 8.001 (d, 1H,5-H), 8.234 (s, 1H, 2′-H), 8.877 (s, 1H, 4-H), 10.547 (s, 1H, CONH),11.103 (br, OH)

Element analysis for: C₁₈H₁₃NO₇Calculated (%): C, 60.85; H, 3.69; N,3.94. Found (%): C, 60.50; H, 3.62; N, 3.64.

Example 11 Synthesis of3-[4′-(5″-methylisooxazol-3″-yl)-amidosulfonyl]phenylamidocarbonyl]-7-methoxy-coumarin (19)

Compound 19 was prepared following the preparation of compound 26,except that 3-carboxy-7-methoxy-coumarin reacted with sulfamethoxazole(SMZ) to give the title compound 19.

Element analysis for: C₂₁H₁₇N₃O₇S.½H₂O Calculated (%): C, 54.31; H,3.91; N, 9.05. Found (%): C, 54.56; H, 3.49; N, 8.90.

Example 12 Synthesis of3-(3′-carboxypropylamidocarbonyl)-7-methoxycoumarin (203)

Compound 203 was prepared following the preparation of compound exceptthat 3-carboxy-7-methoxy-coumarin reacted with γ-amino-butyric acid togive the title compound 203.

¹H-NMR 300 MHz (DMSO): 1.719 (t, 2H, 3′-CH₂), 2.235 (t, 2H, 2′-CH₂),3.311 (t, 2H, 4′-CH₂), 3.861 (s, 3H, 7-OCH₃), 7.001 (d, 1H, 6-H), 7.074(s, 1H, 8-H), 7.861 (d, 1H, 5-H), 8.771 (s, 1H, 4-H)

Element analysis for: C₁₅H₁₅NO₆Calculated (%): C, 59.01; H, 4.95; N,4.59. Found (%): C, 59.05; H, 4.60; N, 4.73.

Example 13 Synthesis of3-[4′-(5″-methylisooxazol-3″)-amidosulfonyl]phenylamidocarbonyl]-7-methoxy-8-methylcoumarin (55)

Compound 55 was prepared following the preparation of compound 26 exceptthat 3-carboxy-7-methoxy-8-methylcoumarin reacted with SMZ to give thetitle compound 55.

Element analysis for: C₂₂H₁₉N₃O₇S Calculated (%): C, 56.28; H, 4.08; N,8.95. Found (%): C, 56.61; H, 4.06; N, 9.01.

Example 14 Synthesis of3-(m-carboxymethylenoxy-phenylamidocarbonyl)-7,8-dimethoxycoumarin (64)

Compound 64 was prepared following the preparation of compound 26 exceptthat 3-carboxy-7,8-dimethoxy-coumarin reacted with m-carboxymethylenoxyaniline to give the title compound 64.

¹H-NMR 300 MHz (DMSO): 3.852 (s, 3H, 8-OCH₃), 3.951 (s, 3H, 7-OCH₃),4.641 (s, 2H, OCH₂), 6.676 (q, 1H, 5′-H), 7.198-7.420 (m, 3H, 4′, 6′,6-H), 7.502 (s, 1H, 2′-H), 7.751 (d, 1H, 5-H), 8.853 (s, 1H, 4-H),10.584 (s, 1H, CONH)

Element analysis for: C₂₀H₁₇NO₈Calculated (%): C, 60.15; H, 4.29; N,3.51. Found (%): C, 60.41; H, 4.65; N, 3.75.

Example 15 Synthesis of3-(4′-guanidinosulfonylphenylamidocarbonyl)-7,8-dimethoxy-coumarin (66)

Compound 66 was prepared following the preparation of compound 26 exceptthat 3-carboxy-7,8-dimethoxy-coumarin reacted with sulfaguanidine (SG)to give the title compound 66.

Element analysis for: C₁₉H₁₈N₄O₇S.2H₂O Calculated (%): C, 47.30; H,4.56; N, 11.61. Found (%): C, 47.34; H, 4.08; N, 11.00.

Example 16 Synthesis of3-(3′-carboxy-4′-hydroxy-phenylamidocarbonyl)-7,8-dimethoxy-coumarin(60)

Compound 60 was prepared following the preparation of compound 26 exceptthat 3-carboxy-7,8-dimethoxy-coumarin reacted with 5-amino-salicylicacid to give the title compound 60.

¹H-NMR 300 MHz (DMSO): 3.849-3.947 (d, 6H, 7,8-bis-OCH₃), 6.962 (d, 1H,5′-H), 7.233 (d, 1H, 6-H), 7.727-7.755 (d, 2H, 5,6′-H), 8.210 (s, 1H,2′-H), 8.813 (s, 1H, 4-H), 10.495 (s, 1H, CONH)

Element analysis for: C₁₉H₁₅NO₈.¼H₂O Calculated (%): C, 58.61; H, 4.01;N, 3.59. Found (%): C, 58.27; H, 3.86; N, 3.92.

Example 17 Synthesis of3-(benzoylhydrazinocarbonyl)-5-methyl-7-methoxycoumarin (210)

Compound 210 was prepared following the preparation of compound 26except that 3-carboxy-5-methyl-7-methoxy-coumarin reacted with benzoylhydrazine to give the title compound 210.

¹H-NMR 300 MHz (DMSO): 2.482 (s, 3H, 5-CH₃), 3.888 (s, 3H, 7-OCH₃),6.979 (d, 2H, 6, 8-H), 7.477-7.583 (q, 2H, 3′,5′-H), 7.500 (t, 1H,5′-H), 7.889 (d, 2H, 2′,6′-H), 8.792 (s, 1H, 4-H), 10.24 (s, 1H, CON),10.868 (s, 1H, CONH)

Example 18 Synthesis of3-(isonicotinoylhydrazinocarbonyl)-5-methyl-7-methoxy coumarin (213)

Compound 213 was prepared following the preparation of compound 26except that 3-carboxy-5-methyl-7-methoxy-coumarin reacted with isoniazidto give the title compound 213.

¹H-NMR 300 MHz (DMSO): 2.553 (s, 3H, 5-CH₃), 3.878 (s, 3H, 7-OCH₃),6.979 (d, 2H, 6, 8-H), 7.935 (d, 2H, 3′,5′-H), 8.781 (s, 1H, 4-He,10.545 (s, 1H, CONH), 11.362 (s, 1H, CONH)

Example 19 Synthesis of3-(3′-carboxy-4′-hydroxy-phenylamidocarbonyl)-5-methyl-7-methoxycoumarin(74)

Compound 74 was prepared following the preparation of compound 26 exceptthat 3-carboxy-5methyl-7-methoxy-coumarin reacted with 5-amino-salicylicacid to give the title compound 74.

Element analysis for: C₁₉H₁₅NO₇Calculated (%): C, 61.79; H, 4.09; N,3.79. Found (%): C, 61.57; H, 4.07; N, 3.81.

Example 20 Synthesis of3-(3′-hydroxy-4′-carboxy-phenylamidocarbonyl)-6-chloro-7-methoxy-coumarin(87)

Compound 87 was prepared following the preparation of compound 26 exceptthat 3-carboxy-6-chloro-7-methoxy-coumarin reacted with4-amino-salicylic acid to give the title compound 87.

¹H-NMR 300 MHz (DMSO): 3.996 (s, 3H, 7-OCH₃), 7.114 (d, 1H, 6′-H), 7.376(s, 1H, 8-H), 7.485 (s, 1H, 2′-H), 7.768 (d, 1H, 5′-H), 8.146 (s, 1H,5-H), 8.839 (s, 1H, 4-H), 10.721 (s, 1H, CONH)

Element analysis for: C₁₈H₁₂ClNO₇Calculated (%): C, 55.47; H, 3.11; N,3.59. Found (%): C, 55.97; H, 3.13; N, 4.48.

Example 21 Synthesis of3-(3′-carboxy-4′-hydroxy-phenylamidocarbonyl)-6-chloro-7-methoxy-coumarin(88)

Compound 88 was prepared following the preparation of compound 26 exceptthat 3-carboxy-6-chloro-7-methoxy-coumarin reacted with5-amino-salicylic acid to give the title compound 88.

¹H-NMR 300 MHz (DMSO): 4.010 (s, 3H, 7-OCH₃), 6.968 (d, 1H, 5′-H), 7.380(s, 1H, 8-H), 7.752 (d, 1H, 6′-H), 8.153 (s, 1H, 5-H), 8.211 (s, 1H,2′-H), 8.817 (s, 1H, 4-H), 10.475 (s, 1H, CONH)

Element analysis for: C₁₈H₁₂ClNO₇Calculated (%): C, 55.47; H, 3.11; N,3.59. Found (%): C, 55.60; H, 3.18; N, 4.1.

Example 22 Synthesis of3-(3′-hydroxy-4′-carboxy-phenylamidocarbonyl)-6-bromo-7-methoxy-coumarin(96)

Compound 96 was prepared following the preparation of compound 26 exceptthat 3-carboxy-6-bromo-7-methoxy-coumarin reacted with 4-amino-salicylicacid to give the title compound 96.

¹H-NMR 300 MHz (DMSO): 3.996 (s, 3H, 7-OCH₃), 7.118 (d, 1H, 6′-H), 7.343(s, 1H, 8-H), 7.496 (s, 1H, 2′-H), 7.774 (d, 1H, 5′-H), 8.306 (s, 1H,5-H), 8.846 (s, 1H, 4-H), 10.722 (s, 1H, CONH)

Example 23 Synthesis of3-(4′-guanidinosulfonylphenylamidocarbonyl)-6-ethyl-7-methoxy-coumarin(32)

Compound 32 was prepared following the preparation of compound 26 exceptthat 3-carboxy-6-ethyl-7-methoxy-coumarin reacted with SG to give thetitle compound 32.

¹H-NMR 300 MHz (DMSO): 1.148 (t, 3H, ethyl-CH₃), 2.572 (q, 2H,ethy-CH₂), 3.896 (s, 3H, OCH₃), 6.690 (br, 4H, guanidino-H), 7.125 (s,1H, 8-H), 7.709 (s, 1H, 5-H), 7.739 (q, 4H, Ar—H), 8.827 (s, 1H, 4-H),10.841 (s, 1H, CONH)

Element analysis for: C₂₀H₂₀N₄O₆S.¼H₂O Calculated (%): C, 53.55; H,4.60; N, 12.48. Found (%): C, 53.49; H, 4.63; N, 12.40.

Example 24 Synthesis of3-(4′-guanidinosulfonylphenylamidocarbonyl)-6-chloro-7-methoxy-coumarin(92)

Compound 92 was prepared following the preparation of compound 26 exceptthat 3-carboxy-6-chloro-7-methoxy-coumarin reacted with SG to give thetitle compound 92.

¹H-NMR 300 MHz (DMSO): 3.999 (s, 3H, 7-OCH₃), 7.407 (s, 1H, 8-H), 7.776(q, 4H, Ar—H), 8.172 (s, 1H, 5-H), 8.860 (s, 1H, 4-H), 10.787 (s, 1H,CONH)

Element analysis for: C₁₈H₁₅ClN₄O₆S Calculated (%): C, 47.95; H, 3.35;N, 12.43. Found (%): C, 47.54; H, 3.45; N, 12.15.

Example 25 Synthesis of3-(3′-hydroxy-4′-carboxy-phenylamidocarbonyl)-7-methoxy-8-methyl-coumarin(43)

Compound 43 was prepared following the preparation of compound 26 exceptthat 3-carboxy-7-methoxy-8-methyl-coumarin reacted with4-amino-salicylic acid to give the title compound 43.

¹H-NMR 300 MHz (DMSO): 2.215 (s, 3H, 8-CH₃), 3.912 (s, 3H, 7-OCH₃),7.081 (d, 1H, 6′-H), 7.182 (d, 1H, 6-H), 7.612 (s, 1H, 2′-H), 7.747 (d,1H, 5-H), 7.872 (d, 1H, 5′-H), 8.834 (s, 1H, 4-H), 10.813 (s, 1H, CONH)

Element analysis for: C₁₉H₁₅NO₇.½H₂O Calculated (%): C, 60.32; H, 4.26;N, 3.70. Found (%): C, 60.26; H, 4.03; N, 4.14.

Example 26 Synthesis of3-(3′-carboxy-4′-hydroxy-phenylamidocarbonyl)-7-methoxy-8-methyl-coumarin(44)

Compound 44 was prepared following the preparation of compound 26 exceptthat 3-carboxy-7-methoxy-8-methylcoumarin reacted with 5-amino-salicylicacid to give the title compound 44

¹H-NMR 300 MHz (DMSO): 2.209 (s, 3H, 8-CH₃), 3.753 (s, 3H, 7-OCH₃),6.959 (d, 1H, 5′-H), 7.168 (d, 1H, 6-H), 7.723 (d, 1H, 6′-H), 7.848 (d,1H, 5-H), 8.197 (s, 1H, 2′-H), 8.794 (s, 1H, 4-H), 10.504 (s, 1H, CONH)

Element analysis for: C₁₉H₁₅NO₇.½H₂O Calculated (%): C, 60.32; H, 4.26;N, 3.70. Found (%): C, 59.66; H, 3.92; N, 3.81.

Example 27 Synthesis of3-(4′-methoxy-phenylamidocarbonyl)-6-nitro-7-hydroxy-8-methyl-coumarin(146)

160 mg (0.604 mmol) of 3-carboxy-6-nitro-7-methoxy-8-methyl-coumarin and2 ml of thionyl chloride was heated to complete the reaction. Extrathionyl chloride was removed and 74.3 mg (0.604 mmol) of p-anisidine, 1ml of pyridine and 1 ml of DMF were added therein and the so-obtainedmixture was heated to complete the reaction. The reaction mixture wasthen filtered and washed by water, diluted hydro chloride, water andethanol, respectively, dried and purified with glacial acetic acid togive 170 mg of the title compound (146).

¹H-NMR 300 MHz (DMSO): 2.280 (s, 3H, Ar—CH₃), 3.740 (s, 3H, OCH₃), 6.941(d, 2H, 3′,5′-H), 7.621 (d, 2H, 2′,6′-H), 8.673 (s, 1H, 5-H), 8.897 (s,1H, 4-H), 10.374 (s, 1H, CONH)

Compounds 110-203, 225-228 were prepared following the same procedure.

Example 28 Synthesis of3-(4′-guanidinosulfonylphenylamidocarbonyl)-6-nitro-7-methoxy-8-methyl-coumarin(169)

Compound 169 was prepared following the preparation of compound 146except that 3-carboxy-6-nitro-7-methoxy-8-methylcoumarin reacted withSG, and purified with DMF to give the title compound 169.

¹H-NMR 300 MHz (DMSO): 2.382 (s, 3H, 8-CH₃), 3.940 (s, 3H, 7-OCH₃),6.677 (br, 4H, guanidino-H), 7.790 (q, 4H, Ar—H), 8.593 (s, 1H, 5-H),8.903 (s, 1H, 4-H), 10.707 (s, 1H, CONH)

Element analysis for: C₁₉H₁₇N₅O₈S.½H₂O Calculated (%): C, 47.10; H,3.75; N, 14.46. Found (%): C, 47.27; H, 3.73; N, 14.58.

Example 29 Synthesis of3-(4′-carboxy-phenylamidocarbonyl)-6-nitro-7,8-dimethoxy coumarin (110)

Compound 110 was prepared following the preparation of compound 146except that 3-carboxy-6-nitro-7,8-methoxycoumarin reacted withp-amino-benzoic acid to give the title-compound 110.

¹H-NMR 300 MHz (DMSO): 3.99-4.06 (q, 6H, 7,8-bis-OCH₃), 7.82 (d, 2H,J=8.7, Ar—H), 7.9 (d, 2H, J=8.7, Ar—H), 8.15 (s, 1H, 5-H), 9.09 (s, 4-H)10.91 (s, 1H, CONH)

Example 30 Synthesis of3-(3′-carboxy-phenylamidocarbonyl)-6-nitro-7,8-dimethoxy-coumarin (111)

Compound 111 was prepared following the preparation of compound 146except that 3-carboxy-6-nitro-7,8-dimethoxycoumarin reacted withm-amino-benzoic acid to give the title compound 111.

¹H-NMR 300 MHz (DMSO): 3.97-4.05 (q, 6H, 7,8-bis-OCH₃), 7.49 (t, 1H,5′-H), 7.67 (d, 1H, 6′-H), 7.76 (d, 1H, 4′H), 7.93 (s, 1H, 2′-H), 8.32(s, 1H, 5-H), 9.08 (s, 1H, 4-H), 10.66 (s, 1H, CONH)

Example 31 Synthesis of3-[4′-(5″,6″-dimethoxypyrimidine-4″)-amidosulfonylphenylamidocarbonyl]-6-nitro-7,8-dimethoxycoumarin (123)

Compound 123 was prepared following the preparation of compound 146except that 3-carboxy-6-nitro-7,8-methoxycoumarin reacted withsulfadoxine (SDM) to give the title compound 123.

¹H-NMR 300 MHz (DMSO): 3.694 (s, 3H, pyrimidine-OCH₃), 3.894 (s, 3H,8-OCH₃), 4.064 (s, 3H, 7-OCH₃), 7.886-7.996 (q, 4H, Ar—H), 7.974 (s, 1H,2″-H), 8.109 (s, 1H, 5-H), 9.092 (s, 1H, 4-H), 10.791 (s, 1H, CONH),10.947 (br, 1H, SO₂NH)

Example 32 Synthesis of3-(3′-hydroxy-4′-carboxyphenylamidocarbonyl)-6-nitro-7-hydroxy-8-methyl-coumarin(148)

Compound 148 was prepared following the preparation of compound 146except that 3-carboxy-6-nitro-7-hydroxy-8-methylcoumarin reacted with4-aminosalicylic acid to give the title compound 148.

¹H-NMR 300 MHz (DMSO): 2.27 (s, 3H, Ar—CH₃), 7.11 (dd, 1H, J=7.8 Hz, 1.8Hz, 6′-H), 7.498 (d, 1H, J=1.8 Hz, 2′-H), 7.775 (d, 1H, J=7.8, 5′-H),8.65 (s, 1H, 5-H), 8.892 (s, 1H, 4-H), 10.69 (s, 1H, CONH)

Example 33 Synthesis of3-(3′-carboxy-4′-hydroxy-phenylamidocarbonyl)-6-nitro-7-hydroxy-8-methyl-coumarin(149)

Compound 149 was prepared following the preparation of compound 146except that 3-carboxy-6-nitro-7-hydroxy-8-methylcoumarin reacted with5-aminosalicylic acid to give the title compound 149.

¹H-NMR 300 MHz (DMSO): 2.268 (s, 3H, Ar—H), 6.971 (d, 1H, J=8.7 Hz,5′-H), 7.747 (dd, 1H, J=8.7 Hz, 2.7 Hz, 6′-H), 8.208 (d, 1H, J=2.7 Hz,2′-H), 8.658 (s, 1H, 5-H), 8.867 (s, 1H, 4-H), 10.403 (s, 1H, CONH)

Element analysis for: C₁₈H₁₂N₂O₉.½H₂O Calculated (%): C, 52.83; H, 3.22;N, 6.85. Found (%): 52.92, 3.26, 6.99.

Example 34 Synthesis of3-[4′-(2″-pyrimidinylamidosulfonyl)phenylamidocarbonyl]-5-methyl-6,8-dinitro-7-hydroy-coumarin(200)

Compound 200 was prepared following the preparation of compound 146except 3-carboxy-5-methyl-6,8-nitro-7-hydroxycoumarin reacted withsulfadiazine (SD) to give the title compound 200.

¹H-NMR 300 MHz (DMSO): 2.291 (s, 3H, 5-CH₃), 7.025 (t, 1H, 5″-H), 7.884(q, 4H, Ar—H), 8.483 (d, 2H, 4″,6″-H), 8.640 (s, 1H, 4-H), 10.705 (s,1H, CONH)

Example 35 Synthesis of3-(4′-amidosulfonylphenylamidocarbonyl)-5-methyl-6,8-dinitro-7-hydroxy-coumarin(198)

Compound 198 was prepared following the preparation of compound 146except that 3-carboxy-5-methyl-6,8-nitro-7-hydroxycoumarin reacted withsulfanilamide to give the title compound 198.

¹H-NMR 300 MHz (DMSO): 2.254 (s, 3H, 5-CH₃), 7.240 (br, 2H, NH₂), 7.788(q, 4H, Ar—H), 8.666 (s, 1H, 4-H), 10.676 (s, 1H, CONH)

Example 36 Synthesis of3-(2′-thiazolamidosulfonylphenylamidocarbonyl)-5-methyl-6,8-dinitro-7-hydroxy-coumarin(201)

Compound 201 was prepared following the preparation of compound 146except that 3-carboxy-5-methyl-6,8-nitro-7-hydroxycoumarin reacted withsulfathiazole (ST) to give the title compound 201.

¹H-NMR 300 MHz (DMSO): 2.291 (s, 3H, 5-CH₃), 6.802 (d, 1H, thiazole-H),7.225 (d, 1H, thiazolyl-H), 7.737 (q, 4H, Ar—H), 8.651 (s, 1H, 4-H),10.667 (s, 1H, CONH)

Example 37 Synthesis of3-(4′-guanidinosulfonylphenylamidocarbonyl)-5-methyl-6,8-dinitro-7-hydroxy-coumarin(199)

Compound 199 was prepared following the preparation of compound 146except that 3-carboxy-5-methyl-6,8-dinitro-7-hydroxycoumarin reactedwith SG to give the title compound 199.

¹H-NMR 300 MHz (DMSO): 2.293 (s, 3H, 5-CH₃), 6.685 (br, 4H,guanidino-H), 7.746 (q, 4H, Ar—H), 8.657 (s, 1H, 4-H), 10.647 (s, 1H,CONH)

Example 38 Synthesis of3-(2′-phenyl-1′,3′,4′-oxadiazol-5′-yl)-7-methoxy-8-methyl coumarin (216)

295 mg (0.84 mmol) of3-(benzoylhydrazinocarbonyl)-7-methoxy-8-methylcoumarin reacted with 4.6ml phosphorus oxychloride at 100° C. for 5 hours, and the reactionmixture was left to be cool and then poured into ice-water, filtrated,washed with water, and dried. 290 mg of the crude product was obtained,and then the crude product was purified with DMF to give 160 mg of thetitle compound 216.

¹H-NMR 300 MHz (DMSO): 2.252 (s, 3H, 8-CH₃), 3.968 (s, 3H, 7-OCH₃),7.174 (d, 1H, 6-H), 7.634 (m, 3H, Ar′—H), 7.812 (d, 1H, 5-H), 8.088 (m,2H, Ar′—H), 8.874 (s, 1H, 4-H)

Compounds 206, 207, 210-212, 215, 216, 219 and 221 in table 2 wereprepared following the same procedure.

Example 39 Synthesis of3-(2′-phenyl-1′,3′,4′-oxadiazol-5′yl)-7-methoxycoumarin (206)

Compound 206 was prepared following the preparation of compound 216except that 3-(benzoylhydrazinocarbonyl)-7-methoxycoumarin reacted withphosphorus oxychloride to give the title compound 206.

¹H-NMR 300 MHz (DMSO): 3.929 (s, 3H, 7-OCH₃), 7.021 (d, 1H, 6-H), 7.085(s, 1H, 8-H), 7.599-7.668 (m, 3H, Ar—H), 7.871 (d, 1H, 5-H), 8.095 (m,2H, Ar—H), 8.898 (s, 1H, 4-H)

Element analysis for: C₁₈H₁₂N₂O₄Calculated (%): C, 67.49; H, 3.78; N,8.75. Found (%): C, 67.57; H, 3.98; N, 8.41.

Example 40 Synthesis of3-[(2′-pyridyl-4″)-1′,3′,4′-oxadiazol-5′yl]-6-hexyl-7-methoxy coumarin(221)

Compound 221 was prepared following the preparation of compound 216except that 3-(isonicotinoylhydrazinocarbonyl)-6-hexyl-7-methoxycoumarin reacted with phosphorus oxychloride to give the title compound221.

¹H-NMR 300 MHz (DMSO): 0.869 (t, 3H, hezyl-CH₃), 1.240 (br, 6H,hezyl-CH₂), 1.574 (t, 2H, hexyl-CH₂), 2.734 (t, 2H, hezyl-CH₂), 3.959(s, 3H, 7-OCH₃), 7.116 (s, 1H, 8-H), 7.699 (s, 1H, 5-H), 8.070 (br, 2H,pyridyl-H), 8.920 (br, 2H, pyridyl-H), 8.921 (s, 1H, 4-H)

Element analysis for: C₂₃H₂₃N₃O₄.3H₂O Calculated (%): C, 60.12; H, 6.36;N, 9.15. Found (%): C, 59.51; H, 5.51; N, 8.96.

Example 41 Synthesis of4-methyl-7-(4′-ethoxycarbonylphenylamidocarbonyl-methylenoxy)coumarin(255)

60 mg (0.256 mmol) of 4-methyl-7-carboxy-methylenoxycoumarin and 2 ml ofthionyl chloride was heated to complete the reaction. Extra thionylchloride was removed and the residue was dissolved in 5 ml of methylenechloride. 44 mg (0.267 mmol) of ethyl 4-amino-benzoate in 5 ml methylenechloride and 3 ml of pyridine were added therein and the reactionmixture was stirred for 0.5 hours to precipitate the solid and thestirration was continued for additional 1 hour. The product wasfiltrated, washed with methylene chloride, and dried to give 80 mg ofthe title compound (255).

¹H-NMR 300 MHz (DMSO): 1.293 (t, 3H, ester-methyl); 2.389 (s, 3H,4-methyl); 4.269 (q, 2H, ester-CH₂), 4.881 (s, 2H, OCH₂), 6.219 (s, 1H,3-H), 7.018 (d, 1H, 8-H), 7.056 (d, 1H, 6-H), 7.712 (d, 1H, 5-H), 7.760(d, 2H, 2′,6′-H), 7.919 (d, 2H, 3′,5′-H), 10.479 (s, 1H, CONH)

Element analysis for: C₂₁H₁₉NO₆Calculated (%): C, 66.13; H, 5.02; N,3.67. Found (%): C, 66.26; H, 4.91; N, 3.81.

Compounds 250-264 in table 2 were prepared following the same procedure.

Example 42 Synthesis of4-methyl-7-phenylamidocarbonyl-methylenoxycoumarin (248)

Compound 248 was prepared following the preparation of compound 255except that ethyl 4-amino-benzoate was replaced by aniline to give thetitle compound 248.

¹H-NMR 300 MHz (DMSO): 2.377 (s, 3H, 4-CH₃), 4.825 (s, 2H, 7OCH₂), 6.208(s, 1H, 3-H), 6.997 (m, 3H, 4′, 6, 8-H), 7.306 (t, 2H, 3′, 5′-H), 7.593(d, 2H, 2′,6′-H), 7.711 (d, 1H, 5-H), 10, 144 (s, CONH)

Element analysis for: C₁₈H₁₅NO₄Calculated (%): C, 69.89; H, 4.89; N,4.53. Found (%): C, 69.61; H, 4.891; N, 4.58.

Example 43 Synthesis of4-methyl-7-(4′-carboxyphenylamidocarbonyl-methylenoxy) coumarin (252)

Compound 252 was prepared following the preparation of compound 255except that ethyl 4-amino-benzoate was replaced by p-amino-benzoic acidto give the title compound 252.

¹H-NMR 300 MHz (DMSO): 2.404 (s, 3H, 4-CH₃), 4.899 (s, 2H, 7-OCH₂),6.235 (s, 1H, 3-H), 7.036 (s, 1H8-H), 7.073 (d, 1H, 6-H), 7.713 (d, 1H,5-H), 7.739-7.924 (q, 4H, Ar—H), 10.491 (s, 1H, CONH)

Element analysis for: C₁₉H₁₅NO₆.¼H₂O Calculated (%): C, 63.77; H, 4.37;N, 3.92. Found (%): C, 63.76; H, 4.28; N, 4.24.

Example 44 Synthesis of4-methyl-7-(4′-hydroxyphenylamidocarbonyl-methylenoxy) coumarin (249)

Compound 249 was prepared following the preparation of compound 255except that ethyl 4-amino-benzoate was replaced by p-amino-phenol togive the title compound 249.

¹H-NMR 300 MHz (DMSO): 2.084 (s, 3H, 4-CH₃), 4.781 (s, 2H, 7-OCH₂),6.230 (s, 1H, 3-H), 6.705-7.390 (q, 4H, Ar—H), 7.014 (s, 1H, 8-H), 7.060(d, 1H, 6-H), 7.723 (d, 1H, 5-H), 9.905 (s, 1H, CONH)

Element analysis for: C₁₈H₁₅NO₅Calculated (%): C, 66.45; H, 4.65; N,4.31. Found (%): C, 66.14; H, 4.62; N, 4.32.

Example 45 Synthesis of4-methyl-7-(3′-carboxy-4′-hydroxyphenylamidocarbonyl-methylenoxy)coumarin(261)

Compound 261 was prepared following the preparation of compound 255except that ethyl 4-amino-benzoate was replaced by 5-amino-salicylicacid to give the title compound 261.

¹H-NMR 300 MHz (DMSO): 2.495 (s, 3H, 4-CH₃), 4.818 (s, 2H, 7-OCH₂),6.233 (s, 1H, 3-H), 6.940 (d, 1H, 6-H), 7.052 (s, 1H, 8-H), 7.077 (d,1H, 5′-H),

Element analysis for: C₁₉H₁₅NO₇ Calculated (%): C, 61.79; H, 4.09; N,3.79. Found (%): C, 61.49; H, 3.96; N, 3.86.

Example 46 Synthesis of4-methyl-7-(3′-trifluoromethylphenylamidocarbonyl-methylenoxy)coumarin(257)

Compound 257 was prepared following the preparation of compound 255except that ethyl 4-amino-benzoate was replaced by 3-fluoromethylaniline to give the title compound 257.

¹H-NMR 300 MHz (DMSO): 2.389 (s, 3H, 4-CH₃), 4.872 (s, 2H, 7-OCH₂),6.220 (s, 1H, 3-H), 7.027-7.075 (m, 2H, 6, 8-H), 7.429 (d, 1H, 6′-H),7.567 (t, 1H, 5′-H), 7.719 (d, 1H, 5-H), 7.857 (d, 1H, 4′-H), 8.096 (s,1H, 2′-H), 10.446 (s, 1H, CONH)

Element analysis for: C₁₉H₁₄F₃NO₄Calculated (%): C, 60.48; H, 3.74; N,3.71. Found (%): C, 60.17; H, 3.45; N, 3.79.

Example 47 Synthesis of4-methyl-7-(3′-trifluoromethyl-4′-nitrophenylamidocarbonylmethylenoxy)coumarin (258)

Compound 258 was prepared following the preparation of compound 255except that ethyl 4-amino-benzoate was replaced by3-fluoromethyl-4′-nitro-aniline to give the title compound 258.

¹H-NMR 300 MHz (DMSO): 2.409 (s, 3H, 4-CH₃), 4.955 (s, 2H, 7-OCH₂),6.243 (s, 1H, 3-H), 7.061 (s, 1H, 8-H), 7.086 (d, 1H, 6-H), 7.734 (d,1H, 5′-H), 8.127 (d, 1H, 6′-H), 8.215 (d, 1H, 5-H), 8.331 (s, 1H, 2′-H),10.945 (s, 1H, CONH)

Element analysis for: C₁₉H₁₃F₃N₂O₆.½H₂O Calculated (%): C, 52.91; H,3.27; N, 6.50. Found (%): C, 53.19; H, 3.05; N, 6.76.

Example 48 Synthesis of4,8-dimethyl-7-(3′-trifluoromethylphenylamidocarbonyl-methylenoxy)coumarin(262)

Compound 262 was prepared following the preparation of compound 255except that 4,8-dimethyl-7-carboxy-methylenoxycoumarin reacted with3-fluoromethylaniline to give the title compound 262.

¹H-NMR 300 MHz (DMSO): 2.291 (s, 3H, 8-CH₃), 2.392 (s, 3H, 4-CH₃), 4.934(s, 2H, 7-OCH₂), 6.237 (s, 1H, 3-H), 7.002 (d, 1H, 6-H), 7.440 (d, 1H,6′-H), 7.564 (d, 1H, 5′-H), 7.603 (d, 1H, 5-H), 7.816 (d, 1H, 4′-H),8.103 (s, 1H, 2′-H), 10.503 (s, 1H, CONH)

Element analysis for: C₂₀H₁₆F₃NO₄Calculated (%): C, 61.38; H, 4.12; N,3.58. Found (%): C, 61.16; H, 4.03; N, 3.67.

Example 49 Synthesis of4,8-dimethyl-7-(3′-hydroxy-4-carboxyphenylamidocarbonylmethylenoxy)-coumarin (264)

Compound 264 was prepared following the preparation of compound 255except that 4,8-dimethyl-7-carboxy-methylenoxycoumarin reacted with4-amino-salicylic acid to give the title compound 264.

¹H-NMR 300 MHz (DMSO): 2.270 (s, 3H, 8-CH₃), 2.371 (s, 3H, 4-CH₃), 4.931(s, 2H, 7-OCH₂), 6.215 (s, 1H, 3-H), 6.958 (d, 1H, 6-H), 7.087 (d, 1H,6′-H), 7.337 (s, 1H, 2′-H), 7.546 (d, 1H, 5′-H), 7.717 (d, 1H, 5-H),10.455 (s, 1H, CONH)

Element analysis for: C₂₀H₁₇NO₇ Calculated (%): C, 62.66; H, 4.47; N,3.65. Found (%): C, 62.43; H, 4.43; N, 3.88.

Example 50 Synthesis of 6-(4′-ethyloxycarbophenylamidocarbonyl)coumarin(265)

A mixture of 95 mg (0.5 mmol) of 6-carboxycoumarin and phosphorouspentachloride in 50 ml toluene was refluxed for 1 hour and the reactionmixture was concentrated. To the residue obtained, 83 mg (0.5 mmol) ofethyl p-amino benzoate and 1 ml of pyridine were added and the refluxwas continued for additional 10 minutes. The reaction mixture was cooleddown and acidified with hydrochloric acid to obtain a solid, which waspurified with ethanol to give 100 mg of the title compound 265

¹H-NMR 300 MHz (DMSO): 1.31 (t, 3H, ester-CH₃), 4.28 (q, 2H, ester-CH₂),6.59 (d, 1H, 3-H), 7.55 (d, 1H, 8-H), 7.92 (d, 2H, Ar′—H), 7.96 (d, 2H,Ar′—H), 8.16 (m, 2H, 4, 7-H), 8.34 (d, 1H, 5-H), 10.68 (s, 1H, CONH)

Element analysis for: C₁₉H₁₅NO₅.½H₂O Calculated (%): C, 65.80; H, 4.65;N, 4.04. Found (%): C, 66.07; H, 4.59; N, 4.06.

Compound 266 was prepared following the same procedure.

Pharmacologic Experiments

Example 1 TGF-β-Induced Cell Growth Inhibition of the Test Compounds onMink Pulmonary Epithelial Cells

Mink pulmonary epithelial cells were seeded in 24 well-plate at adensity of 3×10⁴ cells/well and cultured in modified Eagle's medium(MEM) containing 10% fetal bovine serum in 37° C. and 5% CO₂. Next daythe serum was replaced by a MEM containing 0.2% fetal bovine serum.After 24 hours, the medium was replaced with fresh medium containing 10pmol/L TGF-β and test compounds, and incubated for 24 h. [³H]Thymidinewas added to the medium 2 hours before the termination of incubation.After removing the medium, cells were washed with PBS, dissolved in 0.5mol/L NaOH, and radioactivity was measured. The inhibitory effects oftest compounds are represented as the percentage of Thymidine uptakerecovery (Table 3).

Tab. 3 TGF-β-Induced Cell Growth Inhibition of Tested Compounds in MinkLung Epithelial Cells

TABLE 3 TGF-β-induced cell growth inhibition of tested compounds in MinkLung epithelial cells No. of tested compounds(10 μg/ml) 26 92 73 7 2Inhibition recovery rate 70.7 95.0 15.1 67.1 27.1 on cell growth(%)

Example 2 TGF-β Receptor Binding Antagonism Assay of Test Compounds

Balb/c 3T3 cells were seeded in 24 well-plate and cultured in Dulbecco'smodified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum,in 37° C. and 5% CO₂, for 2-4 days. When cells were at a near-confluent,medium was changed to the binding buffer (50 mmol/L HEPES containingNaCl, KCl, MgSO₄ and CaCl₂). The assay was initiated by addition of 50pmol/L [¹²⁵I]TGF-β1 and test compounds. After incubation for 210-240minutes, the medium was removed, and cells were washed with ice-coldbinding buffer. Non-specific bindings in the presence of 10 nmol/LTGF-β1 were determined. The cells were then solubilized using buffercontaining Triton X-100 and the radioactivity was measured (see Table4).

TABLE 4 TGF-β receptor binding antagonism of tested compounds in Balb/c3T3 cells Compounds Inhibition IC₅₀ Compounds Inhibition IC₅₀ (10 μg/ml)Rate (%) (μg/ml) (10 μg/ml) Rate (%) μg/ml) 1 32.2 42 4.1 2 74.1 13.8 4915.5 3 11.7 55 52.3 6 −6.0 66 52.3 7 94.2 7.8 67 16.2 9 11.4 73 60.0 125.1 79 16.2 14 35.9 83 21.2 21 10.1 87 91.1 22 37.4 88 111.2 5.3 25 11.691 34.7 26 95.4 8.5 92 106.4 27 77.2 93 29.7 31 29.2 96 82.3 33 32.1 10442.8 34 36.4 206 −0.7 37 41.4

Example 3 Protection of Tested Compound Against Chronic Renal FailureCaused by 5/6 Nephrectomy in Rats

The model on chronic renal failure induced by partial renal ablation inrats was set up according to the Guidelines of Pre-clinic Research forNew Drugs.

Male Wistar rats with initial body weights of ˜200 g, anesthetized with35 mg/kg of pentobarbital i.p., the right kidney was removed, and theupper and lower pole parenchymas of the left kidney were resected, stopbleeding, closed the abdominal and sutured. Four weeks followingsurgery, the BUN, creatinine and urinary protein in rats weredetermined. Angiotensin II (AII) level was determined usingradioimmunoassay and TGF-β1 using ELISA methods. Rats were randomlydivided into seven groups, 30/group, with sham, model, Benazepril (4mg/kg/day) and Losartan (10 mg/kg/day) as positive control, and compound149 groups (7.5 mg/kg/day, 15 mg/kg/day and 30 mg/kg/day, respectively).All the groups were administered intragastrically, once per day and 6(six) times per week, till 16 weeks post surgery.

Body weights were weighed to observe the growth of the rats. The aboveindexes were determined every 4 weeks up to the 16^(th) week followingthe initiation of drug administration, and at each time, a number ofanimals were sacrificed and the kidneys were harvested for pathologicalanalysis.

Pathological lesions in chronic renal failure models are mainlyglomerulosclerosis and interstitial fibrosis. According to the damageextents, glomerulosclerosis is divided into five grades (0˜IV). 0 grademeans no glomerulus pathological changes at all, and IV grade meansultimately glomerular sclerosis and glassy pathological changes. 50glomerulus were observed in each animal kidney tissue slide, andpercentage of various grade in each group was calculated based on theabove five grades.

4 weeks after nephrectomy, BUN in serum increased by 111.12% (P<0.01),urinary protein concentration increased by 86.13% (P<0.01), and TGF-β1level increased by 70.84% (P<0.02).

12 weeks after nephrectomy (8 weeks since administration), morphologicalresults demonstrated that the rates of 0 grade glomerylus in residualkidney in 30 mg/kg compound 149 (P<0.05) and Losartan (P<0.05) groupswere higher than that of the model group with significant difference.Glomerulus pathological scores were less than that of the model groupsignificantly. The inflammatory cell infiltration existed in some animalkidney tissues of the Benazepril group was severe, and nephrotubularenlargement as well as protein-like substance also appeared.

16 weeks after nephrectomy (12 weeks since administration), pathologicalresults indicated that the number of grade III glomerulus in the 30mg/kg group of compound 149 and the Losartan group was less than that ofthe model group significantly (P<0.01 and P<0.05, respectively). Theglomerulus pathological score in the positive Benazepril group was thehighest, and the inflammatory cell infiltration in kidney matrix wasmedium-severe, interstitial fibrosis, nephrotubular enlargement as wellas protein-like substance existed.

The results are shown in Tables 5A-E.

TABLE 5 Protection of test compound on chronic renal failure induced by5/6 nephrectomy in rats A. The change of serum creatinine and BUNconcentration in 8^(th) week after administration (following 12 weeksafter nephrectomy) Dose Scr. Change BUN Change Groups (mg/kg) (mg/dL)Rate (%) (mg/dL) Rate (%) Sham — 2.08 ± 0.742 13.00 ± 2.326 125.90↑ Model — 3.06 ± 0.768 47.93↑  29.37 ± 3.079^(#) 28.23↑ Benazepril 4 3.54± 1.140 15.36↑ 37.66 ± 8.895 12.70↓ Losartan 10  2.34 ± 0.268* 23.46↓25.64 ± 5.116 23.89↓ Compound 7.5  2.14 ± 0.500* 30.26↓  22.35 ± 3.120*11.33↓ 149 15  1.80 ± 0.550* 41.34↓ 26.04 ± 4.234  3.98↑ 30 1.89 ± 0.18438.20↓  30.54 ± 11.697 B. The change of serum TGF-β1, Angiotensin II andurinary protein in 8^(th) week after administration (following 12 wkafter nephrectomy) Dose TGF-β1 Change AII Change UP Change Groups(mg/kg) (ng/ml) (%) (pg/ml) (%) (mg/day) (%) Sham 20.1 ± 6.2  54.5 ±22.7 12.7 18.3 ± 2.5  Model 46.33 ± 14.74 130.5↑  94.5 ± 7.4^(#) 73.4↑ 40.7 ± 14.2^(#) 122.5↑  Benazepril 4 40.9 ± 26.6 11.72↓  74.3 ± 13.221.4↓ 51.1 ± 23.6  25.8↑ Losartan 10 18.7 ± 9.2   59.6↓ 96.7 ± 32.1 2.2↑32.7 ± 10.3  19.6↓ Compound 7.5 20.0 ± 6.7   56.8↓  63.9 ± 13.2* 32.4↓30.1 ± 16.6  26.0↓ 149 15 18.6 ± 12.2  59.9↓  49.9 ± 21.3* 47.2↓ 30.4 ±16.2  25.3↓ 30 18.9 ± 10.1  59.2↓  41.0 ± 12.5* 56.6↓ 34.3 ± 12.1  15.7↓C. The change of serum creatinine (Scr.) and BUN concentration in12^(th) week after administration (following 16 weeks after nephrectomy)Dose Scr. Change BUN Change Groups (mg/kg) (mg/dL) Rate (%) (mg/dL) Rate(%) Sham — 2.25 ± 0.39 21.24 ± 3.354 Model —  2.71 ± 0.49^(#)  38.93 ±8.755^(#) 83.32↑  Benazepril 4 2.28 ± 0.70 20.70↑ 39.48 ± 7.109 1.41↑Losartan 10  2.21 ± 0.48* 37.84 ± 5.672 2.80↓ Com. 149 7.5 2.73 ± 0.7819.01↑ 39.42 ± 4.686 1.25↑ 15 2.63 ± 0.38 37.32 ± 5.467 4.14↓ 30  2.10 ±0.71* 22.73↓ 36.60 ± 5.422 5.99↓  0.75↓  2.87↓ 28.82↓ D. The change ofserum TGF-β1, Angiotensin II and urinary protein in 12^(th) week afteradministration (following 16 wk after nephrectomy) Dose TGF-β1 ChangeAII Change UP Change Groups (mg/kg) (ng/ml) (%) (pg/ml) (%) (mg/day) (%)Sham 18.2 ± 8.9 30.0 ± 7.6  16.5 ± 17.3 Model 12.8 ± 7.9 61.7 ± 24.3105.7↑   54.2 ± 26.1^(#)  228↑ Benazepril 4  12.8 ± 14.8 0.57↑ 47.8 ±12.0 22.6↓ 66.3 ± 31.9 22.3↑ Losartan. 10  11.8 ± 12.6 7.48↓  38.9 ±17.4* 37.2↑ 39.3 ± 14.2 18.4↓ Com. 149 7.5 13.6 ± 7.1 6.28↑ 48.3 ± 48.521.6↓ 66.7 ± 38.8 23.1↑ 15 12.3 ± 7.7 3.91↓ 41.3 ± 28.4 33.0↓ 52.3 ±34.4 0.06↓ 30 11.6 ± 6.7 9.38↓  19.2 ± 9.19* 68.6↓ 48.2 ± 31.6 11.1↓ E.Pathological results Glomerulosclerosis Grade (%) Total wk Groups 0 I IIIII IV Grade 8 wk Model 10.0 ± 17.3 38.7 ± 21.2 31.5 ± 17.3 13.6 ± 18.3 1.8 ± 3.3  4.9 ± 1.5 after Benazepril 12.2 ± 19.0 29.6 ± 23.9 28.1 ±18.1 19.6 ± 24.2  5.9 ± 11.2 5.5 ± 2.6 ad. Losartan  54.4 ± 29.2** 31.9± 20.0 13.7 ± 14.0  1.1 ± 3.3** 0  2.8 ± 1.0** Com.149  30 mg/kg 28.3 ±20.8 34.2 ± 23.0  7.5 ± 8.8* 17.5 ± 30.7  6.7 ± 13.4 4.9 ± 3.5  15 mg/kg 37.5 ± 29.3* 27.5 ± 16.3 18.8 ± 9.9  13.3 ± 20.7  2.9 ± 8.2   3.9 ±1.7* 7.5 mg/kg 15.7 ± 19.0 24.0 ± 21.1 30.0 ± 20.8 23.2 ± 28.5  8.0 ±13.0 5.7 ± 2.9 12 wk Model 0 3.3 ± 6.4 29.0 ± 23.4 50.5 ± 18.7 17.6 ±14.1 8.5 ± 1.3 after Benazepril 0 0 19.1 ± 27.1 46.2 ± 15.6 34.8 ± 29.99.8 ± 1.4 ad. Losartan 0 10.0 ± 2.9  45.7 ± 17.7 41.9 ± 25.6  2.8 ± 4.8  7.2 ± 1.3** Com.149  30 mg/kg 0  2.0 ± 4.5   71.3 ± 11.5*  26.7 ± 7.8**0  6.7 ± 0.2**  15 mg/kg 0  8.1 ± 14.1 38.1 ± 27.4 46.2 ± 26.1  9.1 ±12.6 7.7 ± 1.7 7.5 mg/kg 0  2.3 ± 6.3  37.1 ± 16.0 51.4 ± 8.6  13.8 ±20.6 8.6 ± 1.9 Note: *P < 0.05, compared with the model group; ^(#)P <0.05, compared with the sham group; ↑increase; ↓decrease. **P < 0.01,compared with the model group.

The above various parameters with compound 149 treatment are all betterthan those of the Benazepril group and are equivalent to those of theLosartan group. Moreover, pathological results show that the testcompound had no significant affection with the major organs such asheart, liver, spleen, and lungs.

Example 4 Inhibition of the Test Compound on Kidney TubulointerstitialFibrosis Caused by Unilateral Ureteral Obstructed (UUO) in Rats

Male Wistar rats with initial body weights of 180˜230 g were used.Unilateral ureteral obstruction was performed under pentobarbitalanesthesia (35 mg/kg) and sterile conditions. Via a midline incision,the left ureteral was ligated. Sham surgery was performed by making amidline incision but leaving ureteral intact. Following surgery, ratswere randomly divided into sham, model, Benazepril (4 mg/kg/day) andLosartan (10 mg/kg/day) as positive control, and compound 149 (5mg/kg/day, 10 mg/kg/day and 20 mg/kg/day). Starting 2 day beforesurgery, Benazepril, Losartan and compound 149 were administered for 16days orally. BUN and creatinine in serum were determined in the 11^(th)and 16^(th) day (Table 6) following initiation of Benazepril, Losartanand compound 149 treatment, at which time animals were killed and thekidneys were harvested. Tissues were dissected, weighed, fixed in 10%formaldehyde and embedded in paraffin wax for pathological analysis. The9^(th) day serum BUN and creatinine in model group after surgeryincreased 78.7% (P<0.01) and 20.73% (P<0.05) respectively.

TABLE 6 Inhibition of test compound on kidney tubulointerstitialfibrosis caused by unilateral ureteral obstruction (UUO) in rats ChangeChange Dose Scr. Rate BUN Rate Groups (mg/kg) (mg/kg) (%) (mg/dL) (%)Sham — 1.45 ± 0.44 16.23 ± 2.70 Model —  2.20 ± 0.14^(#) 51.58↑  27.54 ±3.32^(#) 69.73↑ Benazepril  4.0 1.92 ± 0.29 12.50↓  20.99 ± 1.58* 23.78↓Losartan 10.0 2.15 ± 0.51  2.31↓ 23.88 ± 2.94 13.30↓ Com.149  5.0  1.58± 0.49* 28.24↓ 23.71 ± 4.17 13.92↓ 10.0  1.61 ± 0.36* 26.50↓  20.76 ±1.56* 24.61↓ 20.0  1.60 ± 0.14* 27.27↓  20.77 ± 2.04* 24.58↓ Note: *P <0.05, compared with model group; ^(#)P < 0.05, compared with controlgroup; ↑increase, ↓decrease.

In this assay, the improvement of each biochemical index with compound149 treatment are more significant than those of the Losartan group, andequivalent to those of the Benazepril group. There was slight differentin the pathological changes: the inflammatory cell infiltration in theBenazepril group was more significant, and 4/7 of the animals had focalabscess formations in the medulla of kidney, many kidney cell necrosisand inflammatory cells and abscess cells overlapped in the Benazeprilgroup. The inflammatory cell infiltration and tubulointerstitialfibrosis were significantly attenuated in both compound 149 and theLosartan groups. That is to say, compound 149 is better than Benazepriland equivalent with Losartan in the pathological results.

Example 5 Primary Acute Toxicity Test for Test Compound

5 g/kg and 10 g/kg of compound 149 were administered orally to mice onceand observed for 14 days. Body weights of 48 hours after administrationin mice were no different. At 14^(th) day after administration, themouse average body weights in 5 g/kg and 10 g/kg groups increased 7 gand 5 g respectively. There was no any other different for every animaland no death were observed.

Example 6 Ames Test

His⁻ type Salmonella typhimurium TA97, TA98, TA100 and TA102 wereemployed. Concentration of the test compound was 0.5, 5.0, 50.0, 500.0,5000.0 μg/plate. S9 was the microsome component of liver homogenate of arat weighted 200 g. The test compound 149 was tested in the presence orabsence of S9.

According to the Salmonella typhimurium/mammalian microsome enzymemutagenic test method revised by Ames (1983), metabolism activated ornon-metabolism activated plate incorporation assay was conducted oncompound 149, and the strain which passed the assay was seeded to theculture medium and incubated at 37° C. under shaking for 15 hours. 100μl compound solutions with various concentrations were added to 0.1 mlof the culture liquid, and then S9 mixture or phosphate buffer wasadded, and the mixtures were incubated in a 37° C. water bath for 20minutes. After that, 2 ml of upper layer agar was added, mixed well andpoured into a plate with lower layer agar and incubated at 37° C. for 48hours. The number of the colonies in each plate was counted.

The results show that the number of colony formation of SalmonellaTyphimurium TA97, TA98, TA100 and TA102 induced by compound 149 did notincrease. It suggests that compound 149 has no mutagenesis.

1. A compound represented by formula (I)

wherein R³ is CONHR₉, wherein R₉ is un-substituted or mono- ormulti-substituted phenyl wherein the substituent is selected from thegroup consisting of hydroxyl, C₁-C₈ alkoxyl, CF₃, carboxyl,alkyloxycarbonyl, OCH₂CO₂H, NO₂, halogen, SO₃H, SO₂NHR₁₁, wherein R₁₁ isselected from the group consisting of hydrogen, amidino, 2″-thiazolyl,3″-(5″-methylisooxazolyl), 2″-pyrimidinyl,2″-(4″,6″-dimethylpyrimidinyl), and 4″-(5″,6″-dimethoxypyrimidinyl); R₄is hydrogen; R₅ is selected from the group consisting of H and C₁-C₄alkyl; R₆ is selected from the group consisting of H, C₁-C₁₂ alkyl,halogen, NO₂, and CONHR₁₃, wherein R₁₃ is substituted phenyl; R₇ isselected from the group consisting of hydroxyl, C₁-C₄ alkyl and alkoxyl;R₈ is selected from the group consisting of H, C₁-C₄ alkyl or alkoxyl,and NO₂; and when R₅, R₆ and R₈ are H, R₇ is OCH₃; or a pharmaceuticallyacceptable salt or hydrate thereof.
 2. The compound according to claim1, wherein R₃ is CONHR₉, wherein R₉ is selected from n-butyric acid, o-,m-, p-hydroxyphenyl, o-, m-, p-carboxyl-phenyl, o-, m-,p-alkyloxycarbophenyl, methoxylphenyl, 3′-hydroxy-4′-carboxyphenyl,3′-salicylyl, 4′-salicylyl, m-CF₃-phenyl, 3′-CF₃-4′-NO₂-phenyl,3′-carboxy-methylenoxyphenyl, 4′-aminosulfonylphenyl,4′-guanidinosulfonylphenyl, 4′-(2″-thiazolaminosulfonyl)phenyl,4′-(5″-methylisooxazolyl-3″-amino sulfonyl)phenyl,4′-(pyrimidinyl-2″-amino sulfonyl)phenyl,4′-(4″,6″-dimethylpyrimidinyl-2″-amino sulfonyl)phenyl, and4′-(5″,6″-dimethoxypyrimidinyl-4″-amino sulfonyl)phenyl; R₄ is hydrogen;R₅ is selected from the group consisting of H and CH₃; R₆ is selectedfrom the group consisting of H, C₂H₅, n-C₆H₁₃, NO₂, Cl, Br, and CONHR₁₃,wherein R₁₃ is selected from the group consisting of 4-benzoic acid andethyl 4-benzoate; R₇ is selected from the group consisting of H, OH,CH₃, and OCH₃; and R₈ is selected from the group consisting of H, CH₃,OCH₃, and NO₂.
 3. The compound according to claim 1, wherein thecompound of formula I is represented by formula (Ib)

wherein R₄, R₅, R₆, R₇, R₈, are as defined in claim 1, R′₂ is selectedfrom the group consisting of H, OH, and CO₂H, R′₃ is selected from thegroup consisting of H, OH, CO₂H, CF₃, and OCH₂CO₂H, R′₄ is selected fromthe group consisting of H, OH, CO₂H, CO₂Et, iodo, NO₂, OCH₃, SO₃H,SO₂NH₂, SONH(C═NH)NH₂,

and R′₅, and R′₆ are each H.
 4. The compound according to claim 2,wherein R₃, R₄, R₅, R₆, R₇, and R₈ are respectively selected from one ofthe combinations in the group consisting of: R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃; R₃=m-CO₂H-phenylamino carbonyl,R₄═R₅═R₆═R₈═H, R₇═OCH₃; R₃=o-CO₂H-phenylamino carbonyl, R₄═R₅═R₆═R₈═H,R₇═OCH₃; R₃=o-OH-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=m-OH-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=p-OH-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=m-OH-p-CO₂H-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=m-CO₂H-p-OH-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=o-CO₂H-p-I-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=4′-ethoxycarbonylphenylaminocarbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=m-CF₃-p-NO₂-phenylaminocarbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃; R₃=4′-aminosulfonylphenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═R₆═R₈═H,R₇═OCH₃; R₃=4′-(2″-pyrimidinylamino aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=4′-[2″-(4″,6″-dimethylpyrimidinylaminosulfonyl)]phenylamino carbonyl,R₄═R₅═R₆═R₈═H, R₇═OCH₃; R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=4′-(5″-methyl-isooxazol-3″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₆═R₈═H, R₇═OCH₃; R₃=p-OCH₃-phenylamino carbonyl, R₄═R₅═R₆═R₈═H,R₇═OCH₃; R₃=p-SO₃H-phenylamino carbonyl, R₄═R₅═R₆═R₈═H, R₇═OCH₃;R₃=p-CO₂H-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=m-CO₂H-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=o-CO₂H-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=p-OH-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=m-OH-p-CO₂H-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=m-CO₂H-p-OH-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=4′-ethoxycarbonylphenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=m-CF₃-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=m-CF₃-4-NO₂-phenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=4′-amino sulfonylphenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=4′-guanidinosulfonylphenylamino carbonyl, R₄═R₅═R₈═H, R₆═C₂H₅,R₇═OCH₃; R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═R₈═H,R₆═C₂H₅, R₇═OCH₃;R₃=4′-(2″-pyrimidinylaminosulfonyl)phenylaminocarbonyl, R₄═R₅═R₈═H,R₆═C₂H₅, R₇═OCH₃;R₃=4′-(4″,6″-dimethylpyrimidinyl-2′-aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃R₃=4′-(5″-CH₃-isooxazol-3″-aminosulfonyl)phenylaminocarbonyl,R₄═R₅═R₈═H, R₆═C₂H₅, R₇═OCH₃; R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═R₈═H,R₆═C₂H₅, R₇═OCH₃; R₃=p-SO₃H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆═C₂H₅,R₇═OCH₃; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=m-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=o-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=m-CO₂H-p-OH-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=o-CO₂H-p-I-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=m-CF₃-4-NO₂-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=4′-aminosulfonylphenylamino carbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈—CH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃,R₈═CH₃; R₃=4′-(2″-pyrimidinylaminosulfonyl)phenylaminocarbonyl,R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=4′-(4″,6″-dimethylpyrimidinyl-2″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃;R₃=4′-(5″-CH₃-isooxazol-3″-aminosulfonyl)phenylaminocarbonyl,R₄═R₅═R₆═H, R₇═OCH₃, R₈═CH₃; R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═R₆═H,R₇═OCH₃, R₈═CH₃; R₃=p-SO₃H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═OCH₃,R₈═CH₃; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃; R₃=mCO₂H-p-OH-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=m-CF₃-p-NO₂-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=m-HO₂CCH₂O-phenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═R₆═H, R₇═R₈═OCH₃;R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=m-CO₂H-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=o-CO₂H-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=o-OH-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=m-OH-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=p-OH-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=m-CO₂H-p-OH-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=m-CF₃-p-NO₂-phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₆═R₈═H, R₅═CH₃,R₇═OCH₃; R₃=4′-(2″-pyrimidinylamino sulfonyl)phenylaminocarbonyl,R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃; R₃=4′-(4″,6″-dimethylpyrimidinyl-2″-aminosulfonyl)phenylamino carbonyl, R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃;R₃=4′-(5″-CH₃-isooxazol-3″-aminosulfonyl)phenylaminocarbonyl,R₄═R₆═R₈═H, R₅═CH₃, R₇═OCH₃; R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₆═R₈═H,R₅═CH₃, R₇═OCH₃; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Cl,R₇═OCH₃; R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Cl, R₇═OCH₃;R₃=m-CO₂H-p-OH-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Cl, R₇═OCH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Cl, R₇═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=0, R₇═OCH₃;R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Cl, R₇═OCH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Cl, R₇═OCH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═R₈═H, R₆═Cl, R₇═OCH₃; R₃=p-CO₂H-aminocarbonyl, R₄═R₅═R₈═H, R₆=Br,R₇═OCH₃; R₃=o-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=Br, R₇═OCH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=Br, R₇═OCH₃;R₃=o-CO₂H-p-I-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=Br, R₇═OCH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═R₈═H, R₆=Br, R₇═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=Br, R₇ ═OCH₃;R₃=4′-aminosulfonylphenylamino carbonyl, R₄═R₅═R₈═H, R₆=Br, R₇═OCH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=Br, R₇═OCH₃;R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=n-Hex, R₇═OCH₃;R₃=o-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=n-Hex, R₇═OCH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═R₈═H, R=Hex, R₇═OCH₃;R₃=o-CO₂H-p-I-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆=n-Hex, R₇═OCH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Hex, R₇═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Hexyl, R₇═OCH₃;R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Hex, R₇═OCH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═R₈═H, R₆═Hex, R₇ ═OCH₃;R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=m-CO₂H-phenylaminocarbonyl, R₄═R₅, R₆═NO₂, R₇═R₈═OCH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=m-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=o-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃═p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃═m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=m-CO₂H-p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=m-CF₃-p-NO₂-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=4′-aminosulfonylphenylamino carbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=4′-(2″-pyrimidinylaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂,R₇═R₈═OCH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═H, R₆═NO₂, R₇═R₈═OCH₃;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂,R₇═R₈═OCH₃; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH,R₈═NO₂; R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=m-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=o-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=m-CO₂H-p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=m-CF₃— phenylamino carbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH, R₈═NO₂;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH,R₈═NO₂; R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H,R₆═C₂H₅, R₇═OH, R₈═NO₂; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅,R₇═OCH₃, R₈═NO₂; R₃=p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OCH₃,R₈═NO₂; R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OCH₃,R₈═NO₂; R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅, R₇═OH,R₈═NO₂; R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═C₂H₅,R₇═OCH₃; R₈═NO₂; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH,R₈═CH₃; R₃=o-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=m-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=o-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=m-CO₂H-p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃R₃=m-CF₃-p-NO₂-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OH,R₈═CH₃; R₃=4′-(2″-pyrimidinylaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H,R₆═NO₂, R₇═OH, R₈═CH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═H, R₆═NO₂, R₇═OH, R₈═CH₃;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂,R₇═OH, R₈═CH₃; R₃=o-CO₂H-p-I-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂,R₇═OH, R₈═CH₃; R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃,R₈═CH₃; R₃=m-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃═o-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=m-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=o-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=m-OH-p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=m-CF₃-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=m-CF₃-p-NO₂-phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃,R₈═CH₃; R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═NO₂,R₇═OCH₃, R₈═CH₃;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂,R₇═OCH₃, R₈═CH₃; R₃=4′-(2″-pyrimidinylaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H, R₆═NO₂, R₇═OCH₃, R₈═CH₃;R₃=p-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃═m-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=o-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃═CF₃-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=4′-aminosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=4′-(2″-pyrimidinylamino aminosulfonyl)phenylaminocarbonyl, R₄═R₅═H,R₆═R₈═NO₂, R₇═OH;R₃=4′-(5″,6″-dimethoxypyrimidinyl-4″-aminosulfonyl)phenylamino carbonyl,R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=4′-(2″-thiazolaminosulfonyl)phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂,R₇═OH; R₃=o-CO₂H-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OH;R₃=p-OH-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OCH₃;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OCH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═R₈═NO₂, R₇═OCH₃;R₃=p-OCH₃-phenylaminocarbonyl, R₄═R₅═H, R₆═Cl, R₇═OH, R₈═NO₂;R₃=4′-guanidinosulfonylphenylaminocarbonyl, R₄═R₅═H, R₆═Cl, R₇═OH,R₈═NO₂; R₃=m-OH-pCO₂H-phenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═Cl,R₈═NO₂; R₃=p-CO₂H-phenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═R₈═NO₂;R₃=m-CO₂H-phenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═R₈═NO₂;R₃=o-CO₂H-phenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═R₈═NO₂;R₃=p-OCH₃-phenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═R₈═NO₂;R₃=p-ethoxycarbophenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═R₈═NO₂;R₃=p-aminosulfonylphenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH, R₆═R₈═NO₂;and R₃=p-guanidinosulfonylphenylaminocarbonyl, R₄═H, R₅═CH₃, R₇═OH,R₆═R₈═NO₂.
 5. A method for preparing a compound according to claim 1,comprising the steps of condensing the substituted 3-carboxy-, or6-carboxy-coumarin, derivative with a corresponding substituted amine.6. The method according to claim 1, wherein the reactants for theamidation reaction are selected from the group consisting of phosphorustrichloride, phosphorus oxychloride, phosphorus pentachloride, thionylchloride, 1,3-dichyclohexylcarbodiimide, dipyridylcarbonate (2-DPC),1,3-diisopropylcarbodiimide (DIPC), and1-(3-dimethylamino-propyl)-3-ethylcarbodiimide (EDC1) and the catalyticagent used is selected from the group consisting of tert-amines,pyridine, 4-dimethylaminopyridine and pyrrolalkylpyridine; and theorganic solvents used comprise dimethylsulfoxide, dichloromethane,toluene, ethylene glycol dimethyl ether, 1,2-dichloroethane,tetrahydrofuran and N,N-dimethylformamide.
 7. A pharmaceuticalcomprising a pharmaceutically effective dosage of a compound accordingto claim 1 and a pharmaceutically acceptable carrier.
 8. Thepharmaceutical composition according to claim 7 wherein thepharmaceutical composition is a tablet, capsule, pH, injection,sustained-release, controlled-release or targeted preparation; and fineparticle delivery systems.
 9. A method for inhibiting transforminggrowth factor β1 comprising administering an amount of a compoundaccording to claim 1 effective to inhibit transforming growth factor β1receptor.
 10. A method for inhibiting angiotensin II (AngII) receptorconverting enzyme comprising administering an amount of a compoundaccording to claim 1 effective to inhibit angiotensin II (AngII)converting enzyme.
 11. A method for treating a chronic renal disordercomprising administering an effective amount of a compound according toclaim 1 to a subject in need thereof.
 12. A method for treatingcardio-cerebrovascular disease comprising administering an effectiveamount of a compound according to claim 1 to a subject in need thereof.13. A method for treating non-insulin dependent diabetes comprisingadministering an effective amount of a compound according to claim 1 toa subject in need thereof.
 14. The method according to claim 12, whereinthe cardio-cerebrovascular disease is hypertension, cerebral embolism,coronary embolism, myocardial infarction, cerebrovascular accident, orstroke or a sequelae thereof.
 15. A method for treating a tumor andpre-cancerous lesion comprising administering an effective amount of acompound according to claim 8 to a subject in need thereof.
 16. Apharmaceutical comprising a pharmaceutically effective dosage of acompound according to claim 4 and a pharmaceutically acceptable carrier.